Gene single strand specific dna and rna endonucleases. Medical search. Frequent questions

Genome-wide CRISPR-Cas9 knockout screens aim to elucidate the relationship between genotype and phenotype by ablating gene expression on a genome-wide scale and studying the resulting phenotypic alterations. (wikipedia.org)
The approach utilises the CRISPR-Cas9 gene editing system, coupled with libraries of single guide RNAs (sgRNAs), which are designed to target every gene in the genome. (wikipedia.org)
The versatility and programmability of Cas9, coupled with the low noise, high knockout efficiency and minimal off-target effects, have made CRISPR the platform of choice for many researchers engaging in gene targeting and editing. (wikipedia.org)
The clustered regularly interspaced short palindrome repeats (CRISPR)/Cas9 system is a gene-editing technology that can introduce double-strand breaks (DSBs) at a target genomic locus. (wikipedia.org)
By using a single guide RNA (sgRNA), the endonuclease Cas9 can be delivered to a specific DNA sequence where it cleaves the nucleotide chain. (wikipedia.org)
A stable AEG‑1‑deficient HT22 neuronal cell line was constructed using CRISPR/Cas9 gene‑editing technology. (spandidos-publications.com)
In this report, we described the application of CRISPR (clustered regularly interspaced short palindromic repeat)/Cas9 (endonuclease) system for efficient genetic editing in E. tenella , and showed that the CRISPR/Cas9 system mediates site-specific double-strand DNA breaks with a single guide RNA. (biomedcentral.com)
Our results extended the utility of the CRISPR/Cas9-mediated genetic modification system to E. tenella , and opened a new avenue for targeted investigation of gene functions in apicomplexan parasites. (biomedcentral.com)
The CRISPR/Cas9 system introduces site-specific double-strand DNA breaks (DSBs) with endonuclease Cas9 in a target sequence that is homologous to the single guide RNA (sgRNA). (biomedcentral.com)
In this study, we present experimental evidence that the CRISPR/Cas9 system is also useful for targeted gene modification in Eimeria . (biomedcentral.com)
CRISPR/Cas9 mediated target gene DSBs. (biomedcentral.com)
Cas9 (Streptococcus pyogenes CRISPR associated protein 9) is an endonuclease enzyme that, when recruited to a specific DNA sequence by the sgRNA (single guide RNA), introduces a double stranded break into the DNA. (genprice.com)
Cas9-expressing HeLa cells can be transduced or electroporated with sgRNA targeting a gene of interest to quickly generate knock-out cell pools or cell lines. (genprice.com)
Here, we report the first application of single Cas9 nickase (Cas9n) to induce gene insertion at a selected locus in cattle. (biomedcentral.com)
Research studies have reported that an extended secondary DNA on the guide crRNA for Cas12a or a hairpin RNA structure added to the sgRNA for Cas9 increases the efficiency and specificity of gene editing. (biorxiv.org)
Cas9 is an endonuclease that cleaves double-stranded DNA in a sequence- specific manner. (biomedcentral.com)
Cas9 associates with a guide RNA of which the first 20 nucleotides pair with the target DNA. (biomedcentral.com)
In addition to this single guide RNA (sgRNA), Cas9 also needs to recognize a multi-nucleotide region that is adjacent to the 3′ end of the target DNA, which is termed PAM (protospacer adjacent motif). (biomedcentral.com)
Several labs have designed sgRNAs to program Cas9 to cleave different regions of HIV-1 DNA that include either essential viral genes or the viral long terminal repeat (LTR). (biomedcentral.com)
Despite the promising possibility that CRISPR/Cas9 could be used to inactivate or even delete proviral DNA from HIV-1 infected cells, an important unanswered question is whether and how HIV-1 might escape from the programmed CRISPR/Cas9 attack, a topic that is fundamental to attempts aimed at HIV treatment and prevention, including the use of small molecule-based antiretroviral therapy and HIV vaccines. (biomedcentral.com)
Rapid escape was observed when non-conserved HIV-1 sequences were attacked, but it did take longer for HIV-1 to escape from Cas9/sgRNAs that targeted the more conserved viral DNA sequences [ 12 ]. (biomedcentral.com)
The majority of the resistance mutations appeared to cluster at the site at which Cas9 was designed to cleave the viral DNA, even though the sgRNA binding site is much bigger. (biomedcentral.com)
Some mutations abrogate the function of viral DNA and will not be selected, while others will be selected because they are not deleterious to the virus, yet generate resistance to Cas9/sgRNA attack because the target DNA sequence is changed. (biomedcentral.com)
Clustered regularly interspaced palindromic repeats (CRISPR)/Cas9 is a gene-editing technology causing a major upheaval in biomedical research. (bmj.com)
CRISPR/Cas9 is a gene-editing technology which involves two essential components: a guide RNA to match a desired target gene, and Cas9 (CRISPR-associated protein 9)-an endonuclease which causes a double-stranded DNA break, allowing modifications to the genome (see figure 1 ). (bmj.com)
Cas9, one of the associated proteins, is an endonuclease that cuts both strands of DNA. (bmj.com)
Cas9 is directed to its target by a section of RNA. (bmj.com)
In order to cut, a specific sequence of DNA of between 2 and 5 nucleotides (the exact sequence depends upon the bacteria which produces the Cas9) must lie at the 3' end of the guide RNA: this is called the protospacer adjacent motif (PAM). (bmj.com)
It is the latter which allows precise genome editing: the homologous section of DNA with the required sequence change may be delivered with the Cas9 nuclease and sgRNA, theoretically allowing changes as precise as a single base-pair. (bmj.com)
One of the most exciting applications of CRISPR/Cas9 is its potential use to treat genetic disorders caused by single gene mutations. (bmj.com)
The clustered regularly interspaced short palindromic repeats (CRISPR)-associated enzyme Cas9 is an RNA-guided endonuclease that uses RNA-DNA base-pairing to target foreign DNA in bacteria. (nature.com)
Cas9-guide RNA complexes are also effective genome engineering agents in animals and plants. (nature.com)
Here we use single-molecule and bulk biochemical experiments to determine how Cas9-RNA interrogates DNA to find specific cleavage sites. (nature.com)
We show that both binding and cleavage of DNA by Cas9-RNA require recognition of a short trinucleotide protospacer adjacent motif (PAM). (nature.com)
Non-target DNA binding affinity scales with PAM density, and sequences fully complementary to the guide RNA but lacking a nearby PAM are ignored by Cas9-RNA. (nature.com)
These results reveal how Cas9 uses PAM recognition to quickly identify potential target sites while scanning large DNA molecules, and to regulate scission of double-stranded DNA. (nature.com)
Figure 1: DNA curtains assay for target binding by Cas9-RNA. (nature.com)
Figure 2: Cas9-RNA remains bound to cleaved products and localizes to PAM-rich regions during the target search. (nature.com)
Figure 3: Cas9-RNA searches for PAMs and unwinds dsDNA in a directional manner. (nature.com)
RNA-guided human genome engineering via Cas9. (nature.com)
Genome engineering of Drosophila with the CRISPR RNA-guided Cas9 nuclease. (nature.com)
The particles contain a Cas9 gene driven by an EF1a promoter, along with a Puromycin selection marker. (magainin.com)
Non-Homologous End Joining often results in the deletion or insertion of several base pairs at the cut site, which, when resulting in a frameshift, causes the functional inactivation of the targeted gene.Cas9 Lentivirus can be used to generate Cas9 expressing cells in almost any mammalian cell line. (magainin.com)
Cells stably expressing Cas9 can then be transduced or electroporated with sgRNA targeting a gene of interest to quickly generate knock-out cell pools or cell lines.The Cas9 Lentiviruses are replication incompetent, HIV-based VSV-G pseudo-typed lentiviral particles that are ready to be transduced into almost all types of mammalian cells, including primary and non-dividing cells. (magainin.com)
CRISPR-Cas9, an RNA-guided genome editing technology, is revolutionizing cell biology due to the ease and efficiency by which it enables genetic manipulation of mammalian cells. (stemcell.com)
Early attempts to apply CRISPR-Cas9 for genome editing in CD34 + HSPCs used plasmids for Cas9 and guide RNA (gRNA) expression, which resulted in low targeting efficiency and high toxicity. (stemcell.com)
Using a recombinant endonuclease-deficient Cas9-based gene promoter pulldown approach coupled with mass spectrometry, we found that MCUb is upregulated in the T2D heart due to loss of glucose homeostasis regulator nuclear receptor corepressor 2 repression, and chromatin immunoprecipitation assays identified peroxisome proliferator-activated receptor α as a mediator of MCUb gene expression in T2D cardiomyocytes. (diabetesjournals.org)
Furthermore, the adaptation of Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) and CRISPR-Associated protein 9 (CRISPR/Cas9) technology has dramatically improved gene editing efficiency at the single gene level. (portlandpress.com)
CRISPR/Cas9 screens, which have revolutionised human and model organism research, have not yet been implemented in malaria parasites due to the need for more complex CRISPR/Cas9 gene targeting vector libraries. (portlandpress.com)
Moreover, since more than half of Plasmodium genes are required for normal asexual blood-stage reproduction, and cannot be targeted using knockout methods, we discuss how CRISPR/Cas9 could be used to scale up conditional gene knockdown approaches to systematically assign function to essential genes. (portlandpress.com)
It complexes with the tracrRNA to direct the Cas9 protein to cut a specific DNA sequence. (corteva.com)
Small RNA molecule constituting part of a 3-component naturally occurring CRISPR-Cas defense system in bacteria and serving as a link for associating the crRNA with the Cas9 protein. (corteva.com)
Guide RNAs (gRNAs) contain the target-specific sequence for guiding Cas9 protein to a genomic location. (idtdna.com)
We guarantee that predesigned Alt-R CRISPR-Cas9 guide RNAs will provide successful editing at the target site, when delivered as a ribonucleoprotein complex as described in the Alt-R Protocols, using Alt-R CRISPR-Cas9 guide RNAs (crRNA:tracrRNA duplex or sgRNA) and either Alt-R S.p. (idtdna.com)

The SpCas9 expression and sgRNA (specific to eyfp ) production, which were controlled by His4 and EtU6 promoter, were constructed in one plasmid. (biomedcentral.com)
Indeed, when the targeted viral DNA regions were sequenced, mutations were identified that interfered with sgRNA recognition. (biomedcentral.com)
The ArciTect™ sgRNA (single guide RNA) or ArciTect™ crRNA (CRISPR RNA) sequences can be designed using the CRISPR Design Tool once a target locus for editing is identified. (stemcell.com)
We offer 3 gRNA formats: crRNA:tracrRNA duplex, crRNA XT:tracrRNA duplex, and single guide RNA (sgRNA). (idtdna.com)

The ERCC5 protein may also function in other cellular processes, including RNA polymerase II transcription, and transcription-coupled DNA repair. (nih.gov)
Comparison of the strengths of the two dnaA promoters in an in vitro transcription system using methylated and unmethylated DNA templates suggests that the effect of methylation on dnaA2P is probably at the level of RNA polymerase/DNA interaction. (jax.org)
Studies of genes transcribed by RNA polymerase II (Pol II) have shown that open and closed chromatin structures are dynamically regulated through multiple mechanisms, including histone modifications, histone variant incorporation, and DNA methylation (reviewed in reference 17 ). (asm.org)
Genome-wide studies in Saccharomyces cerevisiae , for example, have revealed that promoters of most actively transcribed genes are depleted of nucleosomes and that histone density is inversely proportional to the transcription rate within coding regions, suggesting that nucleosomes are dynamically disassembled and reassembled at each passage of the polymerase ( 16 ). (asm.org)
DNA is a nucleic acid which is capable of duplicating itself via the enzyme known as DNA polymerase. (wikibooks.org)
Fluorescence energy transfer between the primer and the beta subunit of the DNA polymerase III holoenzyme. (colorado.edu)
Noteworthy, was our demonstration that although termination of DNA replication did not require specific DNA sequences, some DNA sequences did promote pausing of DNA replication forks in vivo (and DNA polymerase in vitro), and some sequences, such as thosewithin the termination region for SV40 DNA replication, did promote formation of catenated intertwines during separation of sibling chromosomes. (nih.gov)
The detection limit of the RT-LAMP assay was approximately ten viral RNA copies, lower than that of traditional reverse transcriptase polymerase chain reaction (RT-PCR) 100 RNA copies. (biomedcentral.com)
LAMP relies on autocycling strand-displacement DNA synthesis performed by Bst DNA polymerase. (biomedcentral.com)

In the context of genome-wide LOF screens, the aim is to cause gene disruption and knockout. (wikipedia.org)
Our research provides insights into the mechanism of ongoing transposition in the human genome and informs the engineering of retrotransposon proteins for gene therapy. (bvsalud.org)
The viral genome of positive-strand RNA viruses is infectious, as it produces infectious virus upon introduction into a cell. (bvsalud.org)
In this study, we tested the inactivation of the viral genome after RNA fragmentation or cDNA synthesis, using the Trinidad Donkey and TC-83 strains of VEEV. (bvsalud.org)
It can also replace T7 Endonuclease I for genome editing efficiency assessment. (neb.com)
The main reason for the existence of latently infected cells is that proviral DNA becomes integrated into the cellular genome. (biomedcentral.com)
It makes it possible to correct errors in the genome and turn on or off genes in cells and organisms quickly, cheaply and with relative ease. (bmj.com)
RNA-programmed genome editing in human cells. (nature.com)
One-step generation of mice carrying mutations in multiple genes by CRISPR/Cas-mediated genome engineering. (nature.com)
Genome sequencing gives information about the location of genes and provides evidence for the evolutionary links between organisms. (blogspot.com)
Until recently, however, functional analysis of the DED fungi was hampered by the lack of protocols for efficient gene knockout and by the unavailability of a full genome sequence. (sisef.it)
Here we determined genome-wide changes in DNA methylation in the nervous system in neuropathic pain. (jneurosci.org)
However, genome-wide changes in DNA methylation in the nervous system and their roles in neuropathic pain development remain unclear. (jneurosci.org)
Here we used digital restriction enzyme analysis of methylation to quantitatively determine genome-wide DNA methylation changes caused by nerve injury. (jneurosci.org)
In order to isolate a particular gene from human chromosomal DNA, it would be necessary to isolate a sequence of a few hundred or few thousand basepairs from the entire human genome. (addgene.org)
Digesting the human genome with restriction enzymes would yield about two million DNA fragments, which is far too many to separate from each other for the purposes of isolating one specific DNA sequence. (addgene.org)
Here, we review the arrival of genetic screens in malaria parasites to analyse parasite gene function at a genome-scale and their impact on understanding parasite biology. (portlandpress.com)
An RNA-guided endonuclease that complexes with a tracrRNA and a crRNA to introduce a double-strand break at a specific DNA sequence within a genome. (corteva.com)
Adaptation of the bacterial CRISPR-Cas system for genome editing purposes by combining crRNA and tracrRNA into a single guide RNA molecule. (corteva.com)

That is to say why has evolution favored a DNA world over an RNA world given that the two molecules are so similar structurally? (wikibooks.org)
The proteins involved in homologous recombination have to work around other processes that go on inside the nucleus, such as the transcription of DNA in genes into RNA molecules. (elifesciences.org)
At least in plants, DNA regions that become de novo methylated can be defined by homologous RNA molecules in a process termed RNA-directed DNA methylation (RdDM). (degruyter.com)
Viroids are pathogenic circular, non-coding, single-stranded RNA molecules. (degruyter.com)
Viroids are non-encapsidated, single-stranded (ss), 250-400 nucleotide (nt)-long circular RNA molecules that do not encode proteins (1, 2). (degruyter.com)
Upon inoculation, mature Pospiviroidae RNA molecules enter the plant cell nucleus. (degruyter.com)
Therefore, the overall goal of our work is to discover how DNA replication is regulated both in the large chromosomes of cells and in the "mini-chromosomes" of viruses and small extrachromosomal DNA molecules. (nih.gov)
Techniques in chemistry enable isolation and purification of cellular components, such as DNA, but practically this isolation is only feasible for relatively short DNA molecules. (addgene.org)
Single RNA molecules comprised of both crRNA and tracrRNA sequences. (idtdna.com)

The recruitment of this machinery results in the production of viroid-derived small RNAs (vd-sRNAs) that mediate RNA degradation and DNA methylation of cognate sequences. (degruyter.com)
SNL caused DNA methylation changes at 8% of CpG sites with prevailing hypomethylation outside of CpG islands, in introns, intergenic regions, and repetitive sequences. (jneurosci.org)
Gene sequences from normal and carrier animals were compared using the software, codon code Aligner 4.0.4. (researchgate.net)
Typically plasmids contain the minimum essential DNA sequences for this purpose, which includes a DNA replication origin, an antibiotic-resistance gene, and a region in which exogenous DNA fragments can be inserted. (addgene.org)
A piece of DNA comprising nucleotide sequences that are repeated and separated by sequences from invading viruses that serves as part of a naturally occurring CRISPR-Cas defense system in bacteria to fight invading viruses. (corteva.com)
RT-LAMP employs a set of six specially designed primers that recognize eight distinct sequences of the target, and only when three pairs of specific primers match the target sequence exactly can the gene amplification start. (biomedcentral.com)

SeAgo co-purifies with SSB proteins suggesting that they may also be involved in DNA processing. (nih.gov)
Class 2 CRISPR/Cas (Clustered Regularly Interspaced Short Palindromic Repeats/CRISPR-associated proteins) systems, such as Cas12a (Cpf1, subtype V-A) and Cas13a (C2c2, subtype VI), are capable of nonspecific cleavage of ssDNA (single-stranded DNA) and RNA, respectively, in addition to successful gene editing. (biorxiv.org)
Genes are small blocks of DNA that tell the cell which proteins it should create. (wikibooks.org)
His fields of research comprise RNA-mediated gene silencing processes with a focus on epigenetic phenomena, including studies on RNA-directed DNA methylation, the characterization of virus silencing suppressor proteins, the development of plant bioreactor platforms and viroid research. (degruyter.com)
RNA alternative splicing (AS) is an important post-transcriptional mechanism enabling single genes to produce multiple proteins. (nature.com)
The spliceosome complex, composed of at least 170 proteins and several small nuclear RNAs (snRNAs), is the key structure responsible for splicing in eukaryotes 10 . (nature.com)
Over the next 100 years, many significant discoveries lead to the conclusions that genes encode proteins and reside on chromosomes, which are composed of DNA. (addgene.org)
These findings culminated in the central dogma of molecular biology, that proteins are translated from RNA, which is transcribed from DNA. (addgene.org)
Even though all cells of a multicellular organism have the same set of genes, cells in various tissues produce different sets of proteins. (americascupmagazine.com)
Proteomics may be used to study which proteins are expressed under various conditions within a single cell type or to compare protein expression patterns between different organisms. (americascupmagazine.com)
A threadlike molecule of DNA and the associated proteins that support its structure found in the nucleus of most living cells. (corteva.com)
It is organized into genes that specify the production of proteins. (corteva.com)
The cellular process of assembling amino acids into proteins using the nucleotide sequence of messenger RNA (mRNA, see "RNA") as a set of instructions. (corteva.com)
There are several forms of RNA, including messenger RNA (mRNA) which conveys the genetic information from genes (encoded by nucleotides) to proteins (encoded by amino acids). (corteva.com)

Members of the conserved Argonaute (Ago) protein family provide defence against invading nucleic acids in eukaryotes in the process of RNA interference. (nih.gov)
In eukaryotes, DNA methylation refers to the addition of a methyl group to the fifth atom in the six-atom ring of cytosine residues. (degruyter.com)
Although they can be found in archaea and eukaryotes, they play the most significant biological role in bacteria where they can be passed from one bacterium to another by a type of horizontal gene transfer (conjugation), usually providing a benefit to the host, such as antibiotic resistance. (addgene.org)

When purified from its host cells or from heterologous E. coli , SeAgo is loaded with small guide DNAs whose formation depends on the endonuclease activity of the argonaute protein. (nih.gov)
Our latest issue of NEB Expressions covers lyophilized assays for regulated markets, our new phi29-XT RCA Kit, web tool updates, and solutions for automating DNA assembly, sequencing, mutagenesis, cell-free protein expression and more! (neb.com)
SpCas9 was also tagged with enhanced yellow fluorescent protein (EYFP) gene for visualization of the SpCas9 expression signal in subcellular sites of Eimeria . (biomedcentral.com)
Subsequently, we demonstrate that a single Cas9n-induced single-strand break can stimulate the insertion of the natural resistance-associated macrophage protein-1 ( NRAMP1 ) gene with reduced, but still considerable, off-target effects. (biomedcentral.com)
After a systematic selection of target sites and transgenic colonies, we successfully obtained nine exogenous natural resistance-associated macrophage protein-1 ( NRAMP1 ) gene-inserted cows through the Cas9n strategy. (biomedcentral.com)
Cell-free system -- A mixture of cytoplasmic and/or nuclear components from cells used for in vitro protein synthesis, transcription, DNA replication, or other purposes. (nih.gov)
RNase Inhibitor is an acidic, 52 kDa protein that is a potent non-competitive inhibitor of pancreatic-type ribonucleases such as RNase A, RNase B and RNase C. The enzyme is provided as a fusion of the porcine RNase Inhibitor gene with a proprietary 22.5 kDa protein tag. (qiagen.com)
In contrast to RNAi, CRISPR-based gene manipulation requires a guide RNA and a programmable Cas endonuclease protein. (technologynetworks.com)
From this cleavage and repair process, sequence insertions or deletions (indels) often cause errors in translation, leading to functional gene, and subsequent protein, knockdown. (technologynetworks.com)
Presumably, this process involves the interaction between the host's bromodomain-containing viroid RNA-binding protein 1 (VIRP1) and the viroid's TR domain (6). (degruyter.com)
The protein is produced by a recombinant E. coli strain carrying the recombinant thermolabile uracil DNA glycosylase gene isolated from a marine bacteria. (qiagen.com)
Gene therapy displacement of endogenous MCUb with a dominant-negative MCUb transgene (MCUb W246R/V251E ) in vivo rescued T2D cardiomyocytes from metabolic inflexibility and stimulated cardiac contractile function and adrenergic responsiveness by enhancing phospholamban phosphorylation via protein kinase A. We conclude that MCUb represents one newly discovered molecular effector at the interface of metabolism and cardiac function, and its repression improves the outcome of the chronically stressed diabetic heart. (diabetesjournals.org)
The ataxia telangiectasia Rad3-related (ATR) protein responds to UV damage, whereas the ataxia telangiectasia mutated (ATM) protein responds to double-strand breaks (DSBs) caused by ionizing radiation and radiomimetic compounds. (medscape.com)
A protein that breaks down a polynucleotide chain of DNA by cutting the internal bonds linking the nucleotides. (corteva.com)
The amino acid sequence of a protein is encoded by the nucleotide sequence of a corresponding gene. (corteva.com)

L1 spreads through a mechanism termed target-primed reverse transcription, in which the encoded enzyme (ORF2p) nicks the target DNA to prime reverse transcription of its own or non-self RNAs2. (bvsalud.org)
With sequence alignments and structure modeling to guide mutagenesis, we generated enzyme variants across motifs generally conserved or specific to RT subgroups. (bvsalud.org)
The NEBNext Ultrashear™ FFPE DNA Library Prep Kit incorporates enzymatic fragmentation of FFPE DNA using NEBNext UltraShear, a new enzyme mix optimized for use with FFPE DNA that increases library yields and quality, while improving scalability and ease of use. (neb.com)
The endonuclease activity is effective over a wide range of conditions and the enzyme possesses an exceptionally high specific activity. (sydlabs.com)
One unit of Dr. Nuclease corresponds approximately to the required amount of enzyme to digest 37 μg of DNA in the reaction buffer, 50 mM Tris-HCl, pH 8.0, 1 mM MgCl2, for 30 minutes at 37°C. (sydlabs.com)
This suggests that these mutations are not the result of mistakes by the error-prone viral reverse transcriptase (RT) enzyme but rather represent mutations that are generated by the cellular non-homologous end joining (NHEJ) machinery that repairs broken DNA (Fig. 1 ). (biomedcentral.com)
Each enzyme works only on one specific substance (called the substrate). (nih.gov)
Single-stranded exonuclease was determined in a 50 μL reaction containing 10,000 cpm of a radiolabeled single-stranded DNA substrate and 10 μL of enzyme solution incubated for 4 hours at 37°C. (qiagen.com)
Double-stranded endonuclease activity was determined in a 50 μL reaction containing 0.5 μg of pBR322 DNA and 10 μL of enzyme solution incubated for 4 hours at 37°C. (qiagen.com)
E.coli contamination was evaluated using 5 μL samples of enzyme solution were denatured and screened in a TaqMan qPCR assay for the presence of contaminating E.coli genomic DNA using oligonucleotide primers corresponding to the 16S rRNA locus. (qiagen.com)
The enzyme is produced by a recombinant E. coli strain carrying the T4 RNA Ligase gene from bacteriophage T4. (qiagen.com)
One unit is defined as the amount of enzyme required to ligate 50% of 0.4 μg of an equimolar mix of two single-stranded 23 base RNA oligonucleotides (one 5′-phosphorylated) in 20 μL 1X T4 RNA Ligase Buffer following a 30-minute incubation at 37°C. (qiagen.com)
Dilutions of the enzyme were made in 1X T4 RNA Ligase reaction buffer and added to 20 µL reactions containing 0.4 µg of an equimolar mix of two single-stranded 23 base RNA oligonucleotides (one 5′-phosphorylated) and 1X T4 RNA Ligase Buffer. (qiagen.com)
Double-stranded endonuclease is determined in a 50 µL reaction containing 0.5 µg of plasmid DNA and 10 µL of enzyme solution incubated for 4 hours at 37°C. (qiagen.com)

CRISPR RNA maturation by trans-encoded small RNA and host factor RNase III. (nature.com)
Abbreviation for "CRISPR RNA. (corteva.com)

Similar to MilliporeSigma Benzonase nuclease, a genetically engineered endonuclease from Serratia marcescens, Syd Labs recombinant Dr. Nuclease degrades all forms of DNA and RNA while having no proteolytic activity. (sydlabs.com)
Dr. Nuclease is a recombinant endonuclease produced in E. coli. (sydlabs.com)
This obstacle has been overcome by the field of recombinant DNA technology, which enables the preparation of more managable (i.e., smaller) DNA fragments. (addgene.org)
These tools are essential to the field of recombinant DNA, in which many identical DNA fragments can be generated. (addgene.org)
The combination of a DNA fragment with a plasmid or vector DNA backbone generates a recombinant DNA molecule, which can be used to study DNA fragments of interest, such as genes. (addgene.org)
Plasmids that are used most commonly in the field of recombinant DNA technology have been optimized for their use of studying and manipulating genes. (addgene.org)

Whether you are developing cell and gene therapies or viral vaccines, removing contaminating nucleic acids is an important part of viral production. (sigmaaldrich.com)
After the cells produce virus, they are lysed to release the viral particles, DNA, and RNA. (sigmaaldrich.com)
Whether for developing viral vaccines, viral vectors for vaccine, cell and gene therapy, or oncolytic viruses, Benzonase ® endonuclease removes nucleic acid contamination from the final product to meet regulatory standards. (sigmaaldrich.com)
To learn more about optimizing Benzonase ® endonuclease for your process, see our application note " Optimization Strategy and Process Economics of DNA Digestion in Viral Vector Production for Gene Therapy " and data sheet " Optimization of Benzonase ® endonuclease use in virus purification . (sigmaaldrich.com)
Benzonase ® endonuclease intensifies downstream processes by increasing efficiency of these steps and results in higher recovery of viral particles. (sigmaaldrich.com)
VEEV is a select agent and samples containing viral RNA are subject to additional regulations due to their infectious nature. (bvsalud.org)
9 , 10 In addition, chemically modified Cas12a guided-RNA has also been shown to facilitate improved gene correction in mammalian cells through both viral-and non-viral methods compared to the wild-type guide RNA. (biorxiv.org)
This possibility was confirmed by deep sequencing analysis that showed that a number of the resistance-conferring mutations in the viral escape variants indeed matched the mutations that were introduced into the viral DNA in CD4+ T cells that had been infected by HIV-1 for only 36 h [ 13 ]. (biomedcentral.com)
restrict viral infections by recognising and breaking down DNA of invading viruses. (blogspot.com)
RNA splicing plays important roles in viral replication and virus-host interactions 38 . (nature.com)
DNA replication is the primary event that regulates cellular and viral proliferation. (nih.gov)
Drugs that block DNA replication can arrest the spread of cancer cells and eliminate viral pathogens. (nih.gov)
In the past, our research focused on viral genomes as models for DNA replication in mammalian cell nuclei. (nih.gov)

We show that SeAgo is a DNA-guided nuclease preferentially acting on single-stranded DNA targets, with non-specific guide-independent activity observed for double-stranded substrates. (nih.gov)
programmable DNA nuclease. (nih.gov)
Dr. Nuclease acts as an endonuclease that degrades both DNA and RNA in single-stranded, double-stranded, linear, circular, and super-coiled forms. (sydlabs.com)
Dr. Nuclease eliminates DNA/RNA containments in various products with ultra-speed and efficacy. (sydlabs.com)
Endonuclease G (ENDOG) is a nuclear-encoded mitochondrial-localized nuclease. (mdpi.com)
RNA interference (RNAi) was the primary tool for disrupting gene expression (along with zinc finger nucleases (ZFNs) and transcription activator-like effector nuclease (TALENs)) but newer CRISPR-based technologies, including CRISPR knockout (CRISPRko) and CRISPR interference (CRISPRi) are viable options. (technologynetworks.com)

In mammalian cells, active and silent rDNA promoters are characterized by specific histone modifications, DNA methylation, and positioning of the promoter-bound nucleosome (reviewed in reference 22 ). (asm.org)
This double stranded break is repaired in mammalian cells either through Non-Homologous End Joining or Homologous Recombination. (genprice.com)
RNAi using dsRNA has been found to evoke a double-stranded RNA immune response in mammalian cells which can affect cell viability. (technologynetworks.com)
This toxicity can be avoided by switching to short interfering RNAs (siRNAs), which are truncated RNA duplexes that bypass the aforementioned dsRNA cleavage step, retain targeted gene silencing abilities of dsRNA, and do not trigger the deleterious immune response in mammalian cells. (technologynetworks.com)
Emerging evidence indicate that the mammalian checkpoint kinase ATM induces transcriptional silencing in cis to DNA double-strand breaks (DSBs) through a poorly understood mechanism. (elifesciences.org)
In mammalian cells, inhibiting the transcription of genes around a double-strand DNA break depends on a signaling pathway that is activated whenever DNA damage is detected. (elifesciences.org)
Our current research now focuses on two basic, interrelated questions: (1) How do mammalian cells decide where and when to initiate DNA replication? (nih.gov)
2) What are the requirements for DNA replication and transcription at the beginning of mammalian development? (nih.gov)

The NEBNext UltraExpress™ RNA Library Prep Kit has a 3-hour library prep protocol & creates high-quality directional RNA libraries in a single day when paired with poly(A) mRNA enrichment or rRNA depletion kits. (neb.com)
When the RNA fragments complex with a complementary (target) mRNA sequence and an endogenous silencing complex, mRNA is fragmented, preventing its translation. (technologynetworks.com)
Importantly, RNAi-based knockdown is temporary, and effect duration is contingent upon RNA turnover (both for endogenous mRNA and exogenous RNAi modulators). (technologynetworks.com)
While the first mRNA vaccines against an infectious disease have reached the market ( 1 , 2 ), many other medicinal products with RNA as an active substance (see Table 1 ), either vaccines against non-infectious diseases or non-vaccine drugs, are under development ( 3 - 18 ) or even approved ( 19 - 25 ). (frontiersin.org)
There are currently several products containing RNA on the market, and many are under development, among them mRNA, antisense oligonucleotides (ASOs), small interfering RNAs (siRNAs), small activating RNAs (saRNAs), micro-RNAs (mi-RNAs), RNA aptamers and RNA guides. (frontiersin.org)

It's non-specific and cleaves DNA and RNA into smaller fragments, acting on both single stranded and double stranded nuclei acids as well as linear and circular forms of nucleic acids. (sigmaaldrich.com)
The CRISPR/Cas bacterial immune system cleaves bacteriophage and plasmid DNA. (nature.com)

The template polyadenosine tract is recognized in a sequence-specific manner by five distinct domains. (bvsalud.org)
The nuclear localization sequence (nls) of His4 was introduced into the N-terminal of Streptococcus pyogenes (SpCas9) to target the nucleus of the endonuclease SpCas9. (biomedcentral.com)
In DNA a sequence of three bases, which is called a codon, is responsible for the encoding of a single amino acid. (wikibooks.org)
Competition assays provide evidence that DNA strand separation and RNA-DNA heteroduplex formation initiate at the PAM and proceed directionally towards the distal end of the target sequence. (nature.com)
The guide RNA complexes with the Cas endonuclease, is transported to the nucleus, and creates a DNA double-strand break (DSB) at a site pre-programmed in the targeting sequence of the guide RNA. (technologynetworks.com)
Quantitative PCR and high-throughput methods, such as expressed sequence tag analysis, have been used for measuring gene expression and identifying subsets of elm genes that are differentially expressed in the presence of O. novo-ulmi . (sisef.it)
These single-stranded DNA 'tails' are then free to interact with an intact copy of the same DNA sequence from elsewhere in the cell's nucleus, which is used as a guide when repairing the damage. (elifesciences.org)
The study and comparison of entire genomes, including the complete set of genes and their nucleotide sequence and organization, is called genomics. (americascupmagazine.com)

By the year 2000, RNA interference (RNAi) technology had emerged as a fast, simple, and inexpensive technique for targeted gene knockdown, and was routinely being used to study in vivo gene function in C. elegans. (wikipedia.org)
From RNA interference (RNAi) to CRISPR, there are several methods that researchers can use to manipulate gene function, each with its own strengths and weaknesses. (technologynetworks.com)
While the selective inactivation of Ophiostoma genes by insertional mutagenesis remains a challenge, an alternative approach based on RNA interference is now available for down-regulating the expression of targeted genes. (sisef.it)
Since then, he has been working as a postodoc in Dr. Wassenegger's lab (RLP AgroScience GmbH, AlPlanta) and his main field of interest is RNA-interference (RNAi) and RNA-directed DNA methylation (RdDM) in plants. (degruyter.com)
Since then, she has been working as a postodoc in Dr. Wassenegger's lab (RLP AgroScience GmbH, AlPlanta) and her main field of interest is RNA-interference (RNAi) and viroid biology in plants. (degruyter.com)

the section of RNA which binds to the genomic DNA is 18-20 nucleotides. (bmj.com)
RNAi involves transferring a double-stranded RNA (dsRNA) into a cell where it is cleaved and processed into smaller RNA fragments (21 nucleotides). (technologynetworks.com)
We demonstrated that DNA synthesis occurs discontinuously only on one arm of replication forks (the arm where the direction of synthesis is opposite to the direction of fork movement) through the repeated initiation, synthesis and joining of Okazaki fragments (transient nascent DNA chains of 40 to 300 nucleotides). (nih.gov)
Like DNA, RNA is assembled as a chain of nucleotides, but unlike DNA it is more often found in nature as a single-strand folded onto itself. (corteva.com)

There is no activity shown on RNA substrates, making it usable for RT-PCR. (qiagen.com)
For longer single-stranded RNA substrates, overnight incubation at 16°C can improve yield. (qiagen.com)

A programmable dual-RNA-guided DNA endonuclease in adaptive bacterial immunity. (nature.com)
As a graduate student with Julius Adler, I identified the basal body of the bacterial flagellum, develop methods for its purification, and elucidate its fine structure and specific attachments to the bacterial cell envelope. (nih.gov)
Like the bacterial chromosomal DNA, plasmid DNA is replicated upon cell division, and each daughter cell receives at least one copy of the plasmid. (addgene.org)

Once these nucleic acids become broken down, Benzonase ® endonuclease and small DNA fragments can be easily removed by tangential flow filtration. (sigmaaldrich.com)
Plasmids are fragments of double-stranded DNA that typically carry genes and can replicate independently from chromosomal DNA. (addgene.org)
By the 1970s the combined discoveries of restriction enzymes, DNA ligase, and gel electrophoresis allowed for the ability to move specific fragments of DNA from one context to another, such as from a chromosome to a plasmid. (addgene.org)

Mutations in this gene cause xeroderma pigmentosum complementation group G (XP-G), which is also referred to as xeroderma pigmentosum VII (XP7), a skin disorder characterized by hypersensitivity to UV light and increased susceptibility for skin cancer development following UV exposure. (nih.gov)
Subsequent repair of the DSBs is carried out by the host cell through either non-homologous end-joining pathway (NHEJ), leading to insertion and deletion mutations in the targeted genes, or homologous direct repair (HDR) in the presence of a DNA donor template. (biomedcentral.com)
Whithout mutations we would know very little about inheritance and the existence of genes. (intechopen.com)
To the our best of knowledge, this is the first study to report the existence of homozygous and heterozygous mutations at positions 554, 555 and 559 in exon 4 of SLC35A3 gene in Indian Holstein cattle. (researchgate.net)

Targeted approaches for gene knockdown emerged in the 1980s with techniques such as homologous recombination, trans-cleaving ribozymes, and antisense technologies. (wikipedia.org)
Non-Homologous End Joining often results in the deletion or insertion of several base pairs at the cut site, which, when resulting in a frameshift, causes the functional inactivation of the targeted gene. (genprice.com)
Repair after the DNA cut may occur via two pathways: non-homologous end joining, typically leading to a random insertion/deletion of DNA, or homology directed repair where a homologous piece of DNA is used as a repair template. (bmj.com)
Once detected by the cell, blunt-ended DSBs are repaired by the non-homologous end joining (NHEJ) pathway, which is notorious for adding or removing stretches of DNA during repair. (technologynetworks.com)
Therefore, the conversion of the DSB ends from double-stranded to single-stranded DNA, which is necessary to initiate DSB repair by homologous recombination, is responsible for loss of transcription around a DSB in S. cerevisiae . (elifesciences.org)
Homologous recombination' is one of the main mechanisms used by cells to repair DNA double-strand breaks. (elifesciences.org)

The CRISPR/Cas12a RNA-guided complexes have a tremendous potential for nucleic acid detection due to its ability to indiscriminately cleave ssDNA once bound to a target DNA. (biorxiv.org)

The ssRNA(+), likely transferred into the nucleolus, is finally processed by a type III RNase and a DNA LIGASE into unit-length circular RNAs (8) ( Figure 1 ). (degruyter.com)
The T4 RNA Ligase 1 (10,000 U) is intended for molecular biology applications. (qiagen.com)
T4 RNA Ligase catalyzes the ATP-dependent ligation of single-stranded nucleic acids (RNA or DNA) by joining a 5' phosphoryl-terminated nucleic acid donor to a 3' hydroxyl-terminated nucleic acid acceptor through the formation of a 3'→5' phosphodiester bond (1). (qiagen.com)

Early studies in Caenorhabditis elegans and Drosophila melanogaster saw large-scale, systematic loss of function (LOF) screens performed through saturation mutagenesis, demonstrating the potential of this approach to characterise genetic pathways and identify genes with unique and essential functions. (wikipedia.org)
1 It has already been demonstrated that it can be used to repair defective DNA in mice curing them of genetic disorders, 2 and it has been reported that human embryos can be similarly modified. (bmj.com)
DNA, or deoxyribonucleic acid -- The primary genetic material of all cellular organisms and the DNA viruses. (nih.gov)
One of the major functions of the DNA is storage of the genetic information. (wikibooks.org)
It is significant to note the reasons why DNA is the primary method through which all cells pass along genetic information. (wikibooks.org)
Genetic and molecular analyses have led to the identification, cloning and characterization of a few genes that contribute to parasitic fitness in the pathogens. (sisef.it)
The concept of genes as carriers of phenotypic information was introduced in the early 19th century by Gregor Mendel, who later demonstrated the properties of genetic inheritance in peas. (addgene.org)
Chromosomes carry genetic information in the form of genes. (corteva.com)
DNA encodes the heritable genetic information for an organism, (i.e., genetic instructions used in the growth, development, functioning and reproduction) of all known living organisms and many viruses. (corteva.com)
The complete set of genes or genetic material present in a cell or organism. (corteva.com)

To produce viruses for vaccines and cell and gene therapy, cells are genetically transformed. (sigmaaldrich.com)
However, the Benzonase ® endonuclease Safety Plus Emprove ® Expert is produced from a chemically defined medium and each batch is tested for the absence of adventitious viruses and mycoplasma. (sigmaaldrich.com)
Venezuelan equine encephalitis virus (VEEV) is an Alphavirus in the Togaviridae family of positive-strand RNA viruses. (bvsalud.org)
Therefore, RNA isolated from cells infected with BSL-3 select agent strains of VEEV or other positive-strand viruses must be inactivated before removal from high-containment laboratories. (bvsalud.org)
Our cDNA synthesis method also inactivated the genomic RNA of eastern and western equine encephalitis viruses (EEEV and WEEV). (bvsalud.org)
Overall, this work introduces RNA fragmentation and cDNA synthesis as reliable methods for the inactivation of samples containing the genomes of positive-strand RNA viruses. (bvsalud.org)
When applied to large-scale transcriptomics projects with diverse taxonomic sampling, our new method is expected to rapidly expand our knowledge on RNA splicing mechanisms for a wide range of viruses. (nature.com)
My laboratory has developed new technologies and applied them towards understanding the molecular biology and enzymology of DNA replication in animal cells and viruses (SV40, polyomavirus, papillomavirus, and herpes simplex virus), and at the beginning of animal development (mouse preimplantation embryos and frog eggs). (nih.gov)
Additionally, RNA can be amplified by reverse transcription coupled with LAMP (RT-LAMP) in one step, making it suitable to detect RNA viruses such as hMPV. (biomedcentral.com)

The prefix "deoxy" distinguishes DNA from its close relative RNA (ribonucleic acid). (wikibooks.org)

Astrocyte elevated gene-1 (AEG-1), which is also known as metadherin and lysine-rich carcinoembryonic antigen-related cell adhesion molecule 1 co-isolated, was first observed to be upregulated in brain astrocytes from patients with HIV-induced Alzheimer's disease ( 1 ). (spandidos-publications.com)
Mapping, gene -- Determination of the relative positions of genes on a chromosome (or other DNA molecule) and of the distance, in linkage units or physical units, between them. (nih.gov)
When DNA is not self-replicating it exists in the cell as a double stranded helical molecule with the strands lined up anti-parallel to each other. (wikibooks.org)
In RNA there are two possible OH groups that the molecule can form a phosphodiester bond between, which means that RNA is not forced into the same rigid structure as its deoxy counterpart. (wikibooks.org)
A Small RNA molecule encoded by the CRISPR locus constituting part of a 3-component naturally occurring CRISPR-Cas defense system in bacteria. (corteva.com)

Small CRISPR RNAs guide antiviral defense in prokaryotes. (nature.com)

Inside the cell, enzymes create RNA and DNA by facilitating the reaction of ribose with adenosine. (nih.gov)
Furthermore, double-strand DNA has relatively small grooves where damaging enzymes can't attach, making it more difficult for them to 'attack' the DNA. (wikibooks.org)
Double-stranded RNA , on the other hand, has much larger grooves, and therefore, it is more subject to being broken down by enzymes. (wikibooks.org)
This mechanism requires enzymes to eat away at the end of one of the DNA strands on each side of the double-strand break. (elifesciences.org)

DNA methylation differentially enhances the expression of one of the two E. coli dnaA promoters in vivo and in vitro. (jax.org)
The promoter/regulatory region of the dnaA gene, whose gene product is required for the initiation of DNA replication in Escherichia coli K-12, contains an unusually large number of Dam methylation sites. (jax.org)
DNA methylation remodeling in the DRG occurred early after SNL and persisted for at least 3 weeks. (jneurosci.org)
The DNA methylation changes in the injured DRGs recapitulated developmental reprogramming at the neonatal stage. (jneurosci.org)
Methylation reprogramming was correlated with increased gene expression variability. (jneurosci.org)
DNA methylation reprogramming in the DRG thus contributes to nerve injury-induced chronic pain. (jneurosci.org)
Restoring DNA methylation may represent a new therapeutic approach to treat neuropathic pain. (jneurosci.org)
We showed that nerve injury caused DNA methylation changes at 8% of CpG sites with prevailing hypomethylation outside of CpG islands in the dorsal root ganglion. (jneurosci.org)
Reducing DNA methylation induced pain hypersensitivity, whereas increasing DNA methylation attenuated neuropathic pain. (jneurosci.org)

They include coding mRNAs and non-coding (nc) RNAs among them antisense oligonucleotides (ASOs), small interfering RNAs (siRNAs), micro-RNAs (miRNAs), small activating RNAs (saRNAs), RNA aptamers and RNA guides. (frontiersin.org)

DNA double-strand breaks (DSBs) are particularly dangerous for cells, since their inefficient or inaccurate repair can result in deletions and chromosomal translocations that can lead to cancer and/or severe developmental abnormalities in humans. (elifesciences.org)

Note: If you have protospacer designs of your own, or designs from publications, use our design checker tool to assess their on- and off-targeting potential before ordering guide RNAs that are synthesized with using our Alt-R gRNA modifications. (idtdna.com)

Since the DSB ends undergo nucleolytic degradation (resection) of their 5′-ending strands, we investigated the contribution of resection in this DSB-induced transcriptional inhibition. (elifesciences.org)

RNA is essential in various biological roles in coding, decoding, regulation, and expression of genes. (corteva.com)

The two linked strands are twisted in a double helix. (nih.gov)
It is the absence of this group that allows DNA to form its conventional double helix structure. (wikibooks.org)
The modern double helix structure of DNA was first proposed by Watson and Crick, and the functions of DNA were demonstrated in a series of experiments which will be discussed in the next few sections. (wikibooks.org)
Breaks that form across both strands in a DNA double helix are considered the most dangerous type of DNA damage, and can cause a cell to die or become cancerous if they are not repaired accurately. (elifesciences.org)
DNA typically exists in a double-stranded form of two polynucleotide DNA strands coiled around each other to form a double helix. (corteva.com)

Benzonase ® endonuclease is expressed in E. coli W3110, a mutant strain of K12. (sigmaaldrich.com)
In this paper we report that the expression of the dnaA gene is decreased in Dam- strains of E. coli. (jax.org)

We report cryo-electron microscopy structures of the complete human L1 ORF2p bound to structured template RNAs and initiating cDNA synthesis. (bvsalud.org)
Several retroelement and retroviral reverse transcriptases (RTs) support NTA and also template jumping, by which the RT performs continuous complementary DNA (cDNA) synthesis using physically separate templates. (bvsalud.org)
Low-temperature setup at 50°C of the cDNA synthesis step causes delayed C q values because the newly amplified DNA containing dUTP is cleaved by UDG. (qiagen.com)

Additionally, because RNAi reduces gene expression at the post-transcriptional level by targeting RNA, RNAi-based screens only result in partial and short-term suppression of genes. (wikipedia.org)

The decrease in the expression of dnaA was measured in vivo using a dnaA-lacZ gene fusion. (jax.org)

By removing nucleic acids from the sample, this reduces DNA fouling as DNA binds to the resins and ensures maximal binding capacity for virus during chromatography to get the best yields. (sigmaaldrich.com)
This phenomenon has been efficiently harnessed by SHERLOCK (Specific High-sensitivity Enzymatic Reporter unLOCKing) and DETECTR (DNA Endonuclease Targeted CRISPR Trans Reporter) to reliably detect nucleic acids. (biorxiv.org)
DNA is a long chain of linear polymers containing deoxyribose sugars and their covalently bonded bases known as nucleic acids. (wikibooks.org)
These nucleic acid polymers encode for the all of the materials an organism needs to live in the form of genes. (wikibooks.org)

1998), almost all of the ~19,000 genes in C. elegans had been analysed using RNAi-based knockdown. (wikipedia.org)
Methods employed to knockdown or knockout gene products can answer specific questions about gene function, in addition to helping identify and develop therapeutics. (technologynetworks.com)
Achieving a mechanistic and experimental middle ground between CRISPR-mediated gene knockout, whereby gene expression is reduced at the DNA level, and RNAi, in which target knockdown is not permanent, is CRISPRi. (technologynetworks.com)

For example, sgRNAs are most efficient when targeting the coding regions of genes and not the 5' and 3' UTRs. (wikipedia.org)

Here, we developed a platform with engineered crRNAs and optimized conditions that enabled us to detect DNA, DNA/RNA heteroduplex and methylated DNA with higher sensitivity, achieving a limit of detection of in femtomolar range without any target pre-amplification step. (biorxiv.org)

The bases of each strand bind very specifically, A binds with T and C binds with G no other combination exists at least in DNA. (wikibooks.org)

Although its precise biological function remains unclear, its proximity to mitochondrial DNA (mtDNA) makes it an excellent candidate to participate in mtDNA replication, metabolism and maintenance. (mdpi.com)

Assemblages of multiple genes are organized into chromosomes. (corteva.com)

Housekeeping genes in the FSCN1 - ACTB (F-A) locus (Fig. 1a ) have relatively steady expression levels across various tissues and thus exogenous gene silencing resulting from chromatin inactivation might be avoided. (biomedcentral.com)

As gene-modulating tools, RNAi and CRISPR technologies are often grouped together, despite their disparate modes of action and considerations for effective usage. (technologynetworks.com)

The ongoing annotation of these genomes should spark a new wave of interest in the DED pathosystem, as it should lead to the formal identification of genes modulating parasitic fitness. (sisef.it)
The ∼30 Mb genomes of the Plasmodium parasites that cause malaria each encode ∼5000 genes, but the functions of the majority remain unknown. (portlandpress.com)

Among them, an RNA-binding domain bends the template backbone to allow engagement of an RNA hairpin stem with the L1 ORF2p C-terminal segment. (bvsalud.org)

Editing efficiency can be analyzed using ArciTect™ T7 Endonuclease I Kit ( Catalog #76021 ) or flow cytometry, if the experimental design permits. (stemcell.com)
Colony-forming unit (CFU) assays may also be used to assess editing efficiency by plucking and isolating DNA from individual colonies, followed by PCR analysis. (stemcell.com)

DNA is more chemically stable than RNA because it lacks the hydroxyl group on the 2' carbon. (wikibooks.org)

Read-through transcription exists between this gene and the neighboring upstream BIVM (basic, immunoglobulin-like variable motif containing) gene. (nih.gov)
Regulation of nucleosome density is therefore an important mechanism to enable the transcription machinery to access the DNA. (asm.org)
Here we purified full-length L1 ORF2p and biochemically reconstituted robust target-primed reverse transcription with template RNA and target-site DNA. (bvsalud.org)
Previous research has reported that forming a double-strand break in the DNA reduces the levels of transcription for the genes that surround the break, but it was not clear how this occurred. (elifesciences.org)
Instead, the experiments indicate that it is the resection of the DNA around a double-strand break to form single-stranded tails that inhibits transcription in budding yeast. (elifesciences.org)
One of the next challenges will be to see if the resection process makes any contribution to changes in the transcription of genes that surround a double-strand break in mammals as well. (elifesciences.org)

Benzonase endonuclease is an endonuclease that's normally expressed in the bacterium Serratia marcescens . (sigmaaldrich.com)

Members of the Pospiviroidae family replicate in the nucleus through double-stranded RNA intermediates, attracting the host RNA silencing machinery. (degruyter.com)

We guarantee* our predesigned guide RNAs targeting human or mouse, rat, zebrafish, or nematode genes. (idtdna.com)
For other species, we recommend using our proprietary algorithms that enable the design of custom guide RNAs. (idtdna.com)
For more flexibility in guide RNA ordering, CRISPR Custom Guide RNAs allow a wide range of lengths and modifications to support a variety of CRISPR systems and experiments. (idtdna.com)

Authenticase is a mix of structure-specific nucleases that targets mismatch and indel regions (1-10 bp) on double-stranded DNA. (neb.com)
HR is inefficient in many cell types and thus a site-specific double-strand break (DSB) is often introduced by site-specific homing endonucleases or artificial endonucleases, such as zinc finger and TALE nucleases. (biomedcentral.com)

RNA Sanger sequencing analysis was performed in AEG‑1‑deficient HT22 cells and wild‑type HT22 cells without knockout (KO). (spandidos-publications.com)

The simplest way to do this is to use the Benzonase ® endonuclease. (sigmaaldrich.com)
To learn more about switching from the standard Benzonase ® endonuclease to the Benzonase ® endonuclease Safety Plus Emprove ® Expert, download the data sheet or read our regulatory bulletin . (sigmaaldrich.com)

Indeed, several roles for ENDOG have been hypothesized, including maturation of RNA primers during mtDNA replication, splicing of polycistronic transcripts and mtDNA repair. (mdpi.com)
As a post-doctoral fellow with Paul Berg, I developed a subcellular system that allowed simian virus 40 (SV40) DNA to continue replication in vitro. (nih.gov)
In fact, amplification of genes by over replication of certain regions of DNA is one of the primary mechanisms by which cancer cells become resistant to drug therapy. (nih.gov)
We were among the first to develop and exploit sub-cellular systems that allowed SV40 and polyomavirus to complete DNA replication in vitro. (nih.gov)
Results from our lab, as well as from other labs, led to the identification of all of the various DNA replication intermediates in SV40 replication. (nih.gov)
Of particular importance were our studies on DNA replication forks. (nih.gov)

Anatomy2

Organisms6

Diseases1

Chemicals and Drugs46

Analytical, Diagnostic and Therapeutic Techniques and Equipment10

Phenomena and Processes43

Information Science3

Cas954

SgRNA4

Polymerase9

Genome19

Molecules9

Sequences6

Proteins14

Eukaryotes3

Protein15

Enzyme14

CRISPR RNA2

Recombinant6

Viral13

Nuclease6

Mammalian8

MRNA5

Cleaves2

Sequence8

Interference5

Nucleotides4

Substrates2

Bacterial3

Fragments3

Mutations4

Homologous6

Complexes1

Ligase3

Genetic10

Viruses9

Ribonucleic acid1

Molecule5

Prokaryotes1

Enzymes4

Methylation9

SiRNAs1

DSBs1

Protospacer1

Degradation1

Expression of g1

Double helix5

Coli2

CDNA synthesis3

Post-transcriptional1

Vivo1

Nucleic4

Knockdown3

SgRNAs1

Heteroduplex1

Binds1

Mitochondrial1

Chromosomes1

Inactivation1

RNAi1

Genomes2

Hairpin1

Efficiency2

Chemically1

Transcription6

Serratia1

Replicate1

Guide3

Nucleases2

Knockout1

Benzonase2

Replication6