Author Summary RNA interference is a gene regulatory system in which small RNA molecules turn off genes that have similar sequences to the small RNAs. This has become a powerful tool because a researcher can use RNA interference to turn off any gene of interest in order to test its function. There is great interest in identifying the genes required for the RNA interference pathway, and one approach to identifying such genes has been to use RNA interference to turn off potential RNA interference genes and to ask whether RNA interference still functions when these genes are turned off. The goal of our report is to ask how it is possible for RNA interference to turn itself off, using a mathematical model of the system. The results show that RNA interference cannot turn itself off if the RNA interference pathway is too effective to start with, so that experiments in which RNA interference acts on itself will only work in systems having a low efficiency. The results of our model suggest possible ways to
Home , Papers , [EXPRESS] RNA interference-based functional knockdown of the voltage gated potassium channel Kv7.2 in dorsal root ganglion neurons after in vitro and in vivo gene transfer by adeno-associated virus (AAV) vectors. ...
The African trypanosome, Trypanosoma brucei possesses a large and unique intraflagellar structure called the paraflagellar rod (PFR). The PFR is composed of 2 major proteins, PFRA and PFRC. We have generated an inducible mutant trypanosome cell line (snl-2) that expresses linked inverted copies of a PFRA gene, capable of forming a PFRA double-stranded (ds) RNA. When expression of this dsRNA was induced, new PFRA RNA and PFRA protein quickly disappeared and PFR construction was affected, resulting in cell paralysis. This inducible RNA interference (RNAi) effect was fast-acting, heritable and reversible. It allowed us to demonstrate that PFR proteins are able to enter both mature and growing flagella but appear to concentrate differentially in new flagella because of the construction process. The PFR is constructed by a polar assembly process at the distal end of the flagellum resulting in a stable cytoskeletal structure with low turn-over. The inducible RNAi approach will have widespread ...
Histone modifications influence gene expression in complex ways. The RNA interference (RNAi) machinery can repress transcription by recruiting histone-modifying enzymes to chromatin, although it is not clear whether this is a general mechanism for gene silencing or whether it requires repeated sequences such as long terminal repeats (LTRs). We analyzed the global effects of the Clr3 and Clr6 histone deacetylases, the Clr4 methyltransferase, the zinc finger protein Clr1, and the RNA, proteins Dicer, RdRP, and Argonaute on the transcriptome of Schizosaccharomyces pombe (fission yeast). The clr mutants derepressed similar subsets of genes, many of which also became transcriptionally activated in cells that were exposed to environmental stresses such as nitrogen starvation. Many genes that were repressed by the Clr proteins clustered in extended regions close to the telomeres. Surprisingly few genes were repressed by both the silencing and RNAi machineries, with transcripts from centromeric repeats ...
AMSTERDAM, The Netherlands, December 5, 2012 /PRNewswire/ --. uniQure B.V., a leader in the field of human gene therapy, today announced a non-exclusive cross-licensing agreement with Benitec Biopharma Ltd. (ASX: BLT) giving uniQure access to Benitecs proprietary DNA-directed RNA interference (ddRNAi) technology in Huntingtons disease. In return, uniQure granted Benitec non-exclusive access to the Companys AAV5 delivery technology for the development of a ddRNAi therapy for Hepatitis B.. "The cross-licensing agreement with Benitec fully capitalizes on the strength of our advanced AAV platform and our proven ability to deliver therapeutic genes to target cells with high accuracy and efficacy," says Jörn Aldag, CEO of uniQure. "The agreement with Benitec opens up promising new avenues to develop therapies for high unmet medical needs such as Huntingtons disease. While our current programs focus on delivering fully functioning therapeutic genes to remedy faulty or malfunctioning genes, ...
Goodwin Procter associate Daniel Wilson looks into patenting strategies for a powerful new tool for treating disease as well as for creating models of disease.
This RNA interference assay uses chemiluminescence to quantify RNAi target knockdown. No cell lysis is required; the RNAi assay works on culture media.
This RNA interference assay uses chemiluminescence to quantify RNAi target knockdown. No cell lysis is required; the RNAi assay works on culture media.
Background Neurogenesis in the brain of adult mammals occurs throughout life in two locations: the subventricular zone of the lateral ventricle and the subgranular zone of the dentate gyrus in the hippocampus. RNA interference mechanisms have emerged as critical regulators of neuronal differentiation. However, to date, little is known about its function in adult neurogenesis. Results Here we show that the RNA interference machinery regulates Doublecortin levels and is associated with chromatin in differentiating adult neural progenitors. Deletion of Dicer causes abnormal higher levels of Doublecortin. The microRNA pathway plays an important role in Doublecortin regulation. In particular miRNA-128 overexpression can reduce Doublecortin levels in differentiating adult neural progenitors. Conclusions We conclude that the RNA interference components play an important role, even through chromatin association, in regulating neuron-specific gene expression programs. ...
Hypercapnia, elevated partial pressure of CO2 in blood and tissue, develops in many patients with chronic severe obstructive pulmonary disease and other advanced lung disorders. Patients with advanced disease frequently develop bacterial lung infections, and hypercapnia is a risk factor for mortality in such individuals. We previously demonstrated that hypercapnia suppresses induction of NF-κB-regulated innate immune response genes required for host defense in human, mouse, and Drosophila cells, and it increases mortality from bacterial infections in both mice and Drosophila. However, the molecular mediators of hypercapnic immune suppression are undefined. In this study, we report a genome-wide RNA interference screen in Drosophila S2* cells stimulated with bacterial peptidoglycan. The screen identified 16 genes with human orthologs whose knockdown reduced hypercapnic suppression of the gene encoding the antimicrobial peptide Diptericin (Dipt), but did not increase Dipt mRNA levels in air. In ...
Article A novel and quick method to avoid H|sub|2|/sub|O|sub|2|/sub| interference on COD measurement in Fenton system by Na|sub|2|/sub|SO|sub|3|/sub| reduction and O|sub|2|/sub| oxidation. Hydrogen peroxide interference on chemical oxygen demand (COD...
From the abstract: "Hypercapnia, elevated partial pressure of CO2 in blood and tissue, develops in many patients with chronic severe obstructive pulmonary disease and other advanced lung disorders. Patients with advanced disease frequently develop bacterial lung infections ... We previously demonstrated that hypercapnia suppresses induction of NF-κB-regulated innate immune response genes ... However, the molecular mediators of hypercapnic immune suppression are undefined. In this study, we report a genome-wide RNA interference screen in Drosophila S2* cells stimulated with bacterial peptidoglycan. The screen identified 16 genes with human orthologs whose knockdown reduced hypercapnic suppression of the gene encoding the antimicrobial peptide Diptericin (Dipt), but did not increase Dipt mRNA levels in air. In vivo tests of one of the strongest screen hits, zinc finger homeodomain 2 (Zfh2; mammalian orthologs ZFHX3/ATBF1 and ZFHX4), demonstrate that reducing zfh2 function using a mutation or RNA ...
RNA interference (RNAi) is a post-transcriptional process triggered by the introduction of double-stranded RNA (dsRNA) which leads to gene silencing in a sequence-specific manner. The first evidence that dsRNA could achieve efficient gene silencing through RNAi came from studies on the nematode Caenorhabditis elegans. Further analyses in the fruit fly Drosophila melanogaster have contributed greatly toward understanding the biochemical nature of the RNAi pathway. Long dsRNAs are cleaved by the RNase III family member, Dicer, into 19-23 nucleotides (nt) fragments with 5 phosphorylated ends and 2-nt unpaired and unphosphorylated 3 ends.
In the present study, a genome-wide RNA interference screen was combined with an extensive biochemical analysis and quantitative proteomics to better understand the regulation of the heat-shock response (HSR) upon thermal stress. The usage of an endoribon...
TY - JOUR. T1 - A genome-wide loss-of-function screen identifies SLC26A2 as a novel mediator of TRAIL resistance. AU - Dimberg, Lina Y.. AU - Towers, Christina G.. AU - Behbakht, Kian. AU - Hotz, Taylor J.. AU - Kim, Jihye. AU - Fosmire, Susan. AU - Porter, Christopher C.. AU - Tan, Aik-Choon. AU - Thorburn, Andrew. AU - Ford, Heide L.. PY - 2017/4/1. Y1 - 2017/4/1. N2 - TRAIL is a potent death-inducing ligand that mediates apoptosis through the extrinsic pathway and serves as an important endogenous tumor suppressor mechanism. Because tumor cells are often killed by TRAIL and normal cells are not, drugs that activate the TRAIL pathway have been thought to have potential clinical value. However, to date, most TRAIL-related clinical trials have largely failed due to the tumor cells having intrinsic or acquired resistance to TRAIL-induced apoptosis. Previous studies to identify resistance mechanisms have focused on targeted analysis of the canonical apoptosis pathway and other known regulators of ...
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RNA interference (RNAi) is an incredible revolution in the field of functional genomics, a breakthrough in plant molecular genetics. This technology will generate enormous potential for engineering control of gene expres-sion. The success of managing biotic stress using RNAi technology will prove to be biologically and environmentally safe. It is therapeutic in approach as the resistance induced by RNAi is triggered by ds RNA that results in silencing of specific genes before being translated in a homology dependent manner. Over the time, RNAi is significantly proving it as one of the most promiscent management strategy which eliminates certain risks associated with the development of transgenic plants. This review gives an insight into the probability of management of plant diseases caused by various biotic agents viz. fungi, bacteria and viruses using RNA interference technique and host-pathogen related targeted sites ...
Acute myeloid leukemia (AML) with an NPM1 mutation (NPMc+) has a distinct gene expression signature and displays molecular abnormalities similar to mixed lineage leukemia (MLL), including aberrant expression of the PBX3 and HOXA gene cluster. However, it is unclear if the aberrant expression of PBX3 and HOXA is essential for the survival of NPM1-mutated leukemic cells. Methods: Using the gene expression profiling of TCGA and E-MTAB-3444 datasets, we screened for high co-expression of PBX3 and HOXA9 in NPMc+ leukemia patients. We performed NPMc+ depletion and overexpression experiments to examine aberrant H3K79 methylation through epigenetic regulation. Through RNA interference technology and small-molecule inhibitor treatment, we evaluated the effect of methyl-modified H3K79 on cell survival and explored the possible underlying mechanism. Results: We showed that NPMc+ increased the expression of PBX3 and HOXA9, which are both poor prognosis indicators in AML. High PBX3 and HOXA9 expression was ...
For commonly studied genes, where there is only 1 RNAi line in the VDRC GD or KK collection at present, we aim to add a further functional RNAi line to facilitate verification of phenotypes. We chose to use the short hairpin RNAi technology as it is a simpler and more cost-effective method of creating lines than by using long double-stranded RNA. Short hairpins RNAs (shRNAs), containing a 21bp targeting sequence embedded into a micro-RNA (miR-1) backbone, have been shown to be very effective for gene knockdown in both the germline and somatic tissues (Ni et al., 2011). This technology has been used extensively by the Transgenic RNAi Project (TRiP).. To avoid direct duplication of community resources, the VDRC has collaborated with the TRiP team during shRNA design to ensure that the new VDRC lines are as distinct as possible from the TRiP resource. We have used the WALIUM20 vector (for triggering RNAi in soma and germline) in combination with the attP40 landing site, meaning that both the ...
For commonly studied genes, where there is only 1 RNAi line in the VDRC GD or KK collection at present, we aim to add a further functional RNAi line to facilitate verification of phenotypes. We chose to use the short hairpin RNAi technology as it is a simpler and more cost-effective method of creating lines than by using long double-stranded RNA. Short hairpins RNAs (shRNAs), containing a 21bp targeting sequence embedded into a micro-RNA (miR-1) backbone, have been shown to be very effective for gene knockdown in both the germline and somatic tissues (Ni et al., 2011). This technology has been used extensively by the Transgenic RNAi Project (TRiP).. To avoid direct duplication of community resources, the VDRC has collaborated with the TRiP team during shRNA design to ensure that the new VDRC lines are as distinct as possible from the TRiP resource. We have used the WALIUM20 vector (for triggering RNAi in soma and germline) in combination with the attP40 landing site, meaning that both the ...
Title:Therapy for Dominant Inherited Diseases by Allele-Specific RNA Interference: Successes and Pitfalls. VOLUME: 15 ISSUE: 5. Author(s):Delphine Trochet, Bernard Prudhon, Stéphane Vassilopoulos and Marc Bitoun. Affiliation:Inserm/UPMC UMR_S974, CNRS FRE3617, Institut de Myologie, Paris, France.. Keywords:Allele-specific silencing, Dominant inherited diseases, Pitfalls, RNA interference, Single nucleotide substitution, Gene-based therapy.. Abstract:RNA interference (RNAi) is a conserved mechanism for post-transcriptional gene silencing mediated by messenger RNA (mRNA) degradation. RNAi is commonly induced by synthetic siRNA or shRNA which recognizes the targeted mRNA by base pairing and leads to target-mRNA degradation. RNAi may discriminate between two sequences only differing by one nucleotide conferring a high specificity of RNAi for its target mRNA. This property was used to develop a particular therapeutic strategy called "allele-specific-RNA interference" devoted to silence the mutated ...
The use of small interfering RNA (siRNA) molecules in animals to achieve double-stranded RNA-mediated interference (RNAi) has recently emerged as a powerful method of sequence-specific gene knockdown. As DNA-based expression of short hairpin RNA (shRNA) for RNAi may offer some advantages over chemical and in vitro synthesised siRNA, a number of vectors for expression of shRNA have been developed. These often feature polymerase III (pol. III) promoters of either mouse or human origin. To develop a shRNA expression vector specifically for bovine RNAi applications, we identified and characterised a novel bovine U6 small nuclear RNA (snRNA) promoter from bovine sequence data. This promoter is the putative bovine homologue of the human U6-8 snRNA promoter, and features a number of functional sequence elements that are characteristic of these types of pol. III promoters. A PCR based cloning strategy was used to incorporate this promoter sequence into plasmid vectors along with shRNA sequences for RNAi. The
Article describing an optimized protocol for generating short interfering RNAs (siRNAs) or hairpin siRNAs in vitro using T7 RNA Polymerase and annealed DNA oligonucleotide templates. Two RNA interference studies in different mammalian model systems demonstrate the functionality of the synthesized siRNAs.
Examining the knockdown Once cells have been infected, it will be necessary to remove any contaminating uninfected cells. In general, there are two ways to purify and then to analyze RNAi-mediated gene knockdown in cells: one in which the whole population of infected cells are examined, and in the second approach a selected number of individual clonal cell lines are examined. Determining which strategy to perform depends on the nature of the experiment. If the entire population is to be analyzed, either flow cytometric sorting (when GFP-expressing virus is used) or drug selection (when the virus contains an antibiotic-resistance marker) may be used. If a constitutively-expressing small hairpin RNAi vector is used, it will be important to monitor the viability/growth of the cells throughout the procedure. Some gene knockdowns produce slow-growing or lethal phenotypes. This can be difficult to assess when drug selection of the infected cells is used, in which case a GFP marker may be preferred ...
We have led the way in the development of what has been hailed as a major breakthrough in molecular biology: silencing gene expression by RNA interference (RNAi). CSIROs RNAi gene silencing technology is enabling researchers around the world to protect plants and animals from diseases, and to develop new plant varieties with beneficial attributes.
TY - JOUR. T1 - Gene silencing using a heat-inducible RNAi system in Arabidopsis. AU - Masclaux, Frédéric. AU - Charpenteau, Martine. AU - Takahashi, Taku. AU - Pont-Lezica, Rafael. AU - Galaud, Jean Philippe. PY - 2004/8/20. Y1 - 2004/8/20. N2 - Controlling gene expression during plant development is an efficient tool to explore gene function. In this paper, we describe a gene expression system driven by a heat-shock gene promoter (HSP18.2), to trigger the expression of an intron-containing inverted-repeat. RNA interference became a powerful way for gene functional analysis by reverse genetic approaches. However, constitutive gene silencing cannot be used with genes involved in fundamental processes such as embryo viability. Inducible promoters provide an alternative approach for temporal and spatial gene expression control and we described here a new system, complementary to those using chemical gene inducers. To evaluate the efficiency of this system, RNA corresponding to the phytoene ...
Cholesterol levels in the blood are one of the main risk factors for cardiovascular disease. They are controlled by the amount of cholesterol that cells can take in - thus removing it from the blood - and metabolise. The researchers used RNA interference to test the function of each of the genes within 56 regions previously identified by GWAS as being linked with cardiovascular disease. They selectively decreased their action and measured what, if any, changes this induced in cholesterol metabolism. From this they could deduce which of the genes are most likely to be involved in the onset of the disease.. "This is the first wide-scale RNA interference study that follows up on GWAS. It has proven its potential by narrowing down a large list of candidate genes to the few with an important function that we can now focus on in future in-depth studies," explains Rainer Pepperkok at EMBL, who co-led the study with Heiko Runz at the University of Heidelberg. "In principle, our approach can be applied ...
Three years ago Mark Kay MD PhD published the first results showing...Now with three human RNAi gene therapy trials under way Kays initia... Just like any other new drug it is just going to mean that we need t...In traditional gene therapy the inserted DNA produces a gene to replac...With key genes shut off viruses such as hepatitis B hepatitis C or H...,For,Stanford,scientists,,RNAi,gene,therapy,takes,two,steps,forward,,one,step,back,biological,biology news articles,biology news today,latest biology news,current biology news,biology newsletters
MK-1775 is a potent and selective inhibitor of the WEE1 kinase. As of this publication, it is the only WEE1 inhibitor that the authors are aware of currently undergoing evaluation as an anticancer agent in combination with chemotherapy in early-stage clinical trials (19, 20, 28). Previous studies using MK-1775 have shown its potentiation of DNA damage-based therapeutics by forcing unscheduled mitosis and ultimately resulting in apoptosis or mitotic catastrophe (4, 18, 29-32). However, the potential therapeutic effects of WEE1 inhibition in the absence of chemotherapies have not been widely explored. RNA interference knockdown of WEE1 is known to inhibit proliferation of cancer cell lines (13, 33), and more recently, it was shown that MK-1775 alone can induce apoptosis in sarcoma cell lines treated in vitro (34). Our results similarly highlight a requirement for WEE1 activity to maintain cellular viability and genomic stability. Furthermore, we provide the first demonstration of TGI with MK-1775 ...
F. Leulier, P. S Ribeiro, E. Palmer, T. Tenev, K. Takahashi, D. Robertson, A. Zachariou, F. Pichaud, R. Ueda, and P. Meier (2006) Cell Death Differ, 13(10):1663-74.. ...
How to interpret the nature of biological processes, which, when perturbed, cause certain phenotypes, such as human disease, is a major challenge. The completion of sequencing of many model organisms has made reverse genetic approaches [1] efficient and comprehensive ways to identify causal genes for a given phenotype under investigation. For instance, genome-wide knockout strains are now available for Saccharomyces cerevisiae [2, 3], and diverse high throughput RNA interference knockdown experiments have been performed, or are under development, for higher organisms, including C. elegans [4], D. melanogaster [5] and mammals [6, 7].. Compared to the direct genotype-phenotype correlation observed in the above experiments, what is less obvious is how genetic perturbation leads to the change of phenotypes in the complex of biological systems. That is, we might perceive the cell or organism as a dynamic system composed of interacting functional modules that are defined as discrete entities whose ...
Who says you cant be smart, witty, or say anything of substance in 140 characters or less? Alnylam Pharmaceuticals CEO John Maraganore had a zinger ready
Distinct roles for RDE-1 and RDE-4 during RNA interference in Caenorhabditis elegans.: RNA interference (RNAi) is a cellular defense mechanism that uses double-
Plants and fungi can use conserved RNA interference machinery to regulate each others gene expression-and scientists think they can make use of this phenomenon to create a new generation of pesticides.. 6 Comments. ...
Plants and fungi can use conserved RNA interference machinery to regulate each others gene expression-and scientists think they can make use of this phenomenon to create a new generation of pesticides.. 6 Comments. ...
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Therapeutics based on RNA interference (RNAi) gene silencing represent a potentially revolutionary approach to drug development. To realize its broad potential, however, researcher must overcome the significant challenge posed by the delivery of the RNAi trigger (siRNA) to the target. While progress has been steady, as reflected in a range of pre-clinical animal models, clinical data is only starting to emerge, and proof-of-concept for RNAi gene silencing following systemic administration is yet to be obtained. In a recent publication, researchers discussed findings that nanoparticles of siRNAs formulated in cyclodextrin-containing polycations (RONDEL) successfully trafficked to cancer tissues in patients following systemic intravenous administration and triggered the degradation of the targeted cancer-related gene RRM2. This marks an important milestone in the clinical development of RNAi therapeutics and provides the rationale for the further development of RONDEL siRNA delivery for cancer and ...
High-throughput screening of cellular effects of RNA interference (RNAi) libraries is now being increasingly applied to explore the role of genes in specific cell biological processes and disease states. However, the technology is still limited to specialty laboratories, due to the requirements for robotic infrastructure, access to expensive reagent libraries, expertise in high-throughput screening assay development, standardization, data analysis and applications. In the future, alternative screening platforms will be required to expand functional large-scale experiments to include more RNAi constructs, allow combinatorial loss-of-function analyses (e.g. genegene or gene-drug interaction), gain-of-function screens, multi-parametric phenotypic readouts or comparative analysis of many different cell types. Such comprehensive perturbation of gene networks in cells will require a major increase in the flexibility of the screening platforms, throughput and reduction of costs. As an alternative for ...
Alnylam Pharmaceuticals, Inc. (Nasdaq:ALNY), the leading RNAi therapeutics company, today announced that the Company presented new pre-clinical data highlighting its next generation
The work that led to the discovery and functional characterization of BCL-2 family proteins relied on a variety of cell and molecular biology approaches, genetically engineered mouse models, and RNA interference-based methods. With the synthesis of the first validated BH3 mimetics, ABT-737 and navitoclax (6, 7), came the ability to inhibit BCL-2 and BCL-XL directly with cell-permeable small molecules, facilitating a host of new discoveries. Since its introduction, the BCL-2-selective inhibitor ABT-199/GDC-0199 (venetoclax) has been used to define the role of BCL-2 in specific hematopoietic lineages (32), a variety of hematologic cancers (25, 33-38), and even some solid tumors, including estrogen receptor-positive breast cancer (39). In addition, certain roles for BCL-XL have been deduced using a subtractive parsing method that compares the effects of ABT-737 or navitoclax to those of venetoclax (38, 40-42). For example, Bah and co-workers used ABT-737 and venetoclax to demonstrate that BCL-XL ...
A genome-wide RNAi screen in Drosophila S2 cells identified 41 genes capable of specifically modifying Aβ secretion without affecting general secretion or viability. As expected, the four γ-secretase complex components showed the most potent Aβ-lowering effects upon knockdown, with ,10% Aβ remaining. This validated the RNAi screen because it demonstrated that the approach could robustly identify relevant Aβ-modulator genes in an unbiased screen. There was a large difference in the magnitude of phenotype between these four genes and the other genes identified, suggesting that no other essential γ-secretase complex protein exists as expected (Edbauer et al., 2003), but rather proteins that serve a modulatory role in Aβ generation. Indeed, another high-throughput RNAi screen found a potent modulator of BACE1 activity but did not describe the identification of any γ-secretase modulators (Majercak et al., 2006). Knockdown of pigeon, the Drosophila homolog of the recently identified GSAP ...
For more information, please visit www.dicerna.com. About GalXC™ RNAi Technology Platform. GalXC™ is a proprietary technology platform invented by Dicerna to discover and develop RNAi-based therapies designed to silence disease-driving genes in the liver. Compounds produced via GalXC are intended to be broadly applicable across multiple therapeutic areas involving the liver, including rare diseases, chronic liver diseases, cardiovascular diseases and viral infectious diseases. Using GalXC, Dicerna scientists attach N-acetylgalactosamine sugars directly to the extended region of the Companys proprietary RNAi molecules, yielding multiple proprietary conjugate delivery configurations. Many of the conjugates produced via GalXC incorporate a folded motif known as a tetraloop in the extended region. The tetraloop configuration, which is unique to Dicernas GalXC compounds, allows flexible and efficient conjugation to the targeting ligands, and stabilizes the RNAi duplex which the Company believes ...
RNA inteference (RNAi) refers to a natural phenomenon in which the expression of a double stranded RNA leads to the degradation, through a complex process, of mRNAs to which one of its strands is complementary. Several years ago our lab along with several others, including those of William Hahn (DFCI), Eric Lander (Broad Institute), and Nir Hacohen (MGH), formed a consortium to generate an arrayed library of RNAi reagents that target every human gene and to develop the methodologies for using the library in a high-throughput fashion. This consortium, now called The RNAi Consortium (TRC), has generated, in a lentiviral vector, 5 distinct RNAi constructs for ost human genes. Each construct consists of a short hairpin RNA (shRNA) that directs the expression of a small interfering RNA (siRNA) that in turn causes the degradation of the mRNA which it recognizes. Our goal is to create constructs for every human and mouse gene for a total library size of about 300,000 constructs (5 constructs/gene for ...
microscopy improve the axial resolution of far-field fluorescence microscopy as much as threefold to sevenfold. However, establishing the phase difference of the wave fronts in the sample is a problem yet to be solved. Here we show that the phase difference is encoded in the microscopes transfer of the spatial frequencies that match the distance of the interference peaks. As a result the phase difference is readily extracted through a Fourier transform of the image. Our method is relevant to all microscopes that exploit the interference of counterpropagating waves to improve the axial and the lateral resolution.. © 2002 Optical Society of America. Full Article , PDF Article ...
An estimated $1.8 billion dollars is spent for each new successful drug developed, primarily in failed clinical trials due to lack of efficacy and safety. New approaches for rapid identification and early preclinical validation of novel therapeutic targets are crucial to make important go/no-go decisions and curb the cost of developing new cancer treatments. For decades, genetically engineered mouse models have provided a powerful platform to study disease initiation and maintenance, the tumor microenvironment and the responsiveness of cancers to known or novel therapeutics; however, the long lead times and high costs required to develop, intercross and maintain models with various cancer predisposing gene combinations have limited their practical utility in the drug discovery process. RNA interference (RNAi), a mechanism that controls gene expression, can be exploited experimentally to silence nearly any gene target. By expressing synthetic short hairpin RNAs (shRNAs) in mice, RNAi serves as ...
Sigma-Aldrich offers abstracts and full-text articles by [Ming Ying, Guangfeng Chen, Yu Qiu, Xiujuan Shi, Chen Zhang, Qiuke Wang, Shuzhang Yang, Lixia Lu, Qionglan Yuan, Guotong Xu, Zibing Jin, Qiang Wu, Xiaoqing Liu].
Two leaders in genome editing, MilliporeSigma and The Wellcome Sanger Institute, have joined forces to make the first ever arrayed lentiviral CRISPR knockout libraries for human and mouse genomes.. Genome-wide loss-of-function screening is a powerful approach to discover genes and pathways that underlie biological processes. Now complete knockout is achievable with two optimized gRNAs per gene. Minimized clone number ensures the most specific screening possible while controlling time and cost.. Early screening experiments used RNA interference (RNAi), which enables gene knockdown but not complete gene knockout. Thus, RNAi may miss important gene hits when total loss of gene expression is required. CRISPR-Cas9 technology allows more efficient loss-of-function screening in mammalian cells on a large-scale.. The advent of pooled CRISPR libraries brought efficient genome-wide knockout but required the additional step of deep sequencing deconvolution for accurate hit identification. Arrayed screening ...
chemical synthesized siRNA and rapidly dividing cells - posted in siRNA, microRNA and RNAi: Hi all, I am new to RNAi area. Your suggestion and opinion are very important to me If i am to use some rapidly dividing cells (e.g. HEK 293 cells) to perform RNAi study, is it not wise to use chemically synthesized siRNA? I have this wonder because in this case the RNAi effects may be too insignificant to be detectable. Is my concept correct? since i think for chemically synthesized siRNA, i...
Intradigm Corp., a leading developer of targeted, systemic RNA interference (RNAi) therapeutics, and Agilent Technologies Inc. (NYSE: A) today announced that Intradigm has selected Agilent to manufact...