• Here, we review the structural elements required for designing mRNA vaccine constructs for effective in vitro synthetic transcription reactions. (mdpi.com)
  • The unprecedently speedy development of mRNA vaccines against severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) was enabled with previous innovations in nucleoside modifications during in vitro transcription and lipid nanoparticle delivery materials of mRNA. (mdpi.com)
  • Whereas the in vitro translation efficiency of these two mRNA isoforms was about equal, the mHIF-1alphaI.2 5'-untranslated region (5'-UTR) conferred significantly higher in vivo luciferase reporter gene activity than the mHIF-1alphaI.1 5'-UTR. (uzh.ch)
  • Additionally, the 5′ and 3′ UTRs flanking the coding region regulate mRNA translation, half-life, and subcellular localization [2]. (researchgate.net)
  • 6143)-enables easy cloning of a template DNA into a pre-designed In-Fusion cloning-ready plasmid system for in vitro transcription with the Takara IVTpro T7 mRNA Synthesis Kit. (takarabio.com)
  • 6144)-includes a highly-efficient T7 RNA polymerase optimized for preparing high yields of mRNA through in vitro transcription. (takarabio.com)
  • Takara IVTpro mRNA Synthesis System includes all components necessary from DNA template preparation to high-yield in vitro transcription reaction. (takarabio.com)
  • Cloning Kit for mRNA Template enables easy cloning of a template DNA into a pre-linearized plasmid (included in the kit) for downstream in vitro transcription reactions. (takarabio.com)
  • Takara IVTpro mRNA Synthesis Kit enables the production of large amounts of mRNA through in vitro transcription using an optimized, highly efficient T7 RNA polymerase. (takarabio.com)
  • The kit synthesizes up to 200 μg or more of mRNA per 20 μl reaction, includes DNase I to digest the template DNA after the in vitro transcription reaction and a lithium chloride (LiCl) solution to purify the synthesized mRNA for downstream applications. (takarabio.com)
  • Multiple reports clearly demonstrate that G4s are enriched in mRNA 5? (grantome.com)
  • Interestingly, an alternative initiation codon has been shown to be present within the coding region of p53 mRNA. (iisc.ac.in)
  • In this mechanism, the ribosomes are recruited to a complex RNA structural element known as 'Internal Ribosome Entry Site (IRES)', generally present in the 5' untranslated region (UTR) of mRNA. (iisc.ac.in)
  • The IRES directing the translation of p53 is in the 5'-untranslated region of the mRNA, whereas the IRES mediating the translation of ΔN-p53 extends further into the protein-coding region. (iisc.ac.in)
  • Transfection of in vitro synthesized bicistronic RNAs confirmed the presence of IRES elements within p53 mRNA. (iisc.ac.in)
  • According to the current model, scanning ribosomes translate the 5′-most uORF (uORF1) and, under non-starvation conditions, reinitiate translation at downstream uORFs 2, 3, or 4 and subsequently dissociate from the mRNA, keeping GCN4 translation repressed. (nih.gov)
  • We have also investigated the roles of various eIFs, tRNAi and the 40S subunit in scanning the mRNA 5′ untranslated region and in accurately identifying the AUG initiation codon. (nih.gov)
  • These sequences typically include the 5′-untranslated region in mRNA including the ribosome binding site (Shine-Dalgarno sequence) and in some cases the start codon (Klinkert and Narberhaus 2009 ). (springer.com)
  • Mutations affecting the hunchback mRNA target sites for Nanos-dependent translational repression were found to disrupt translational repression in vivo, but had little effect on Nanos RNA binding in vitro. (embl.de)
  • Ribonucleoprotein complex immunoprecipitation and biotinylated RNA pull-down assays demonstrated that PARP-14 forms a complex with the mRNA-destabilizing protein tristetraprolin (TTP) and a conserved adenylate-uridylate-rich element in the TF mRNA 3′ untranslated region. (imperial.ac.uk)
  • When this conversion occurs in the coding region of mRNA, it results in an altered nucleotide codon and, therefore, can change the amino acid sequence of the coded protein in what is referred to as a re-coding editing event. (biomedcentral.com)
  • [ 9 ] They discovered one low frequency single nucleotide polymorphism (SNP) in the noncoding 5' untranslated (UTR) region of the gene RBM8A in 4 of the cases and another low frequency noncoding SNP in the first intron of the same gene. (medscape.com)
  • Some biologic features plaque assay, we determined that approximately 360 viral of the SARS-CoV described in vivo and in vitro differ genomes were required to generate a PFU. (cdc.gov)
  • This study was conducted to evaluate in vivo and in vitro development of in vitro-matured equine oocytes fertilized by intracytoplasmic sperm injection. (bioone.org)
  • Four of the five stem-loops of the 5' UTR have 70% to 100% conserved sequences across the various members of the genus. (news-medical.net)
  • The pre-linearized plasmid contains the T7 promoter, transcription initiation sequence (AGG), 5'- and 3'-UTRs (untranslated region), and poly(A) sequence. (takarabio.com)
  • Where do you find the 5' and 3' untranslated regions (UTRs)? (chiyuanye.com)
  • 2) containing structurally different 5'-termini which might modulate translation initiation. (uzh.ch)
  • ii) identifying a higher-order assembly of initiation factors 1, 2, 3, and 5-the multifactor complex (MFC)-and elucidating its roles in PIC assembly and accurate start codon selection. (nih.gov)
  • Its innermost region consists of a cone-shaped core that includes two copies of the (positive sense) ssRNA genome, the enzymes reverse transcriptase, integrase and protease, some minor proteins, and the major core protein. (wikipedia.org)
  • The 5′ two-thirds of the genome contains two ORFs (ORF 1a and ORF 1b) that encode 15-16 nonstructural proteins (nsps), and the other one-third of the genome consists largely of genes encoding structural and accessory proteins [ 5 ]. (biomedcentral.com)
  • We show that full-length and C-terminal Nanos proteins bind to RNA in vitro with high affinity, but with little sequence specificity. (embl.de)
  • Poly(adenosine 5′-diphosphate [ADP]-ribose)-polymerase (PARP)-14 belongs to a family of intracellular proteins that generate ADP-ribose posttranslational adducts. (imperial.ac.uk)
  • Yet, the UbV inhibits the essential cleavage activities of the protease in vitro and in cells, and it reduces viral replication in cell culture by almost five orders of magnitude. (bvsalud.org)
  • In some species (e.g., bovine), evidence indicates that decreases in IGFBP-4 and -5 levels in estrogen-dominant preovulatory follicles are likely due, in part, to increased protease activity, whereas lower levels of IGFBP-2 are not due to increased proteolysis. (bioone.org)
  • Increased IGFBP-4 and -5 protease along with lower amounts of IGFBP-4 binding activity and greater amounts of free IGF-I are some of the earliest developmental changes documented in bovine growing antral follicles. (bioone.org)
  • These protease-induced decreases in IGFBP-4 and -5 likely cause increased levels of bioavailable (or free) IGFs that stimulate steroidogenesis and mitogenesis in developing dominant follicles, which ultimately prepare the follicle(s) and oocyte(s) for successful ovulation and fertilization. (bioone.org)
  • Polymorphisms including single nucleotide polymorphisms (SNPs) in genes involved in xenobiotic metabolism and in genes involved in oestrogen biosynthesis and metabolism might affect circulating estrogen levels and modulate the individual susceptibility to environmental carcinogens in relation to developing BC [ 5 ]. (biomedcentral.com)
  • To date, more than 100 genes have been reported as associated with non-syndromic hearing loss [ 5 ]. (biomedcentral.com)
  • In the recent past, the presence and function of a ROSE-like RNA-thermometer located in the 5′UTR of the rhamnosyltransferase genes rhlAB has been reported in wild type P. aeruginosa . (springer.com)
  • For this purpose, the non-pathogenic production host P. putida KT2440 containing the rhlAB genes with the native P. aeruginosa 5′-UTR region was used. (springer.com)
  • A new U.S. study published on the bioRxiv * preprint server reports the use of an amiloride-based scaffold that targets a viral RNA structure known to be essential in viral replication in the 5'-untranslated region (UTR) of the virus. (news-medical.net)
  • RNA G4s are enriched in important regulatory regions of RNAs thus potentially implicating them in different aspects of RNA metabolism. (grantome.com)
  • Here, using tiling arrays and RNAs from wild-type and mutant bacteria grown in vitro, ex vivo and in vivo, we have analysed the transcription of its entire genome. (ox.ac.uk)
  • 500 nucleotides), at least two of which are involved in virulence in mice, we have identified antisense RNAs covering several open-reading frames and long overlapping 5' and 3' untranslated regions. (ox.ac.uk)
  • We sequenced the EDN1 untranslated regions of a small subset of patients with PAH, predicted the effect in silico, and used a luciferase assay with the different genotypes to analyze its influence on gene expression. (stanford.edu)
  • rs397751713) and another in the 3'regulatory region (rs2859338) that altered luciferase activity in vitro depending on their genotype. (stanford.edu)
  • The researchers say this heralds the application of these molecules as chemical probes to explore viral biology and produce specific antivirals targeting RNA regions of the coronavirus genome. (news-medical.net)
  • TRS-L, which is located at the 5′ terminus of the coronavirus genome, shares high sequence identity with TRS-B. TRS-L acts as an acceptor for the complementary TRS-B donor sequence during sgmRNA synthesis [ 5 ] through a similarity-assisted copy-choice mechanism. (biomedcentral.com)
  • As a first step, the 5'UTRs mediating the translation of both p53 and ΔN-p53 were cloned in the intercistronic regions of bicistronic constructs. (iisc.ac.in)
  • First, we will test the hypothesis that protein kinase C activates a trans-acting repressor protein(s), which binds to the 5' or 3' untranslated region of the mature apo E transcript and inhibits translation. (unthsc.edu)
  • These will include the synthesis of modified apo E transcripts, in vitro translation assays, coupled with gel mobility shift assays and UV cross-linking studies. (unthsc.edu)
  • In collaboration with Jon Lorsch's group, we demonstrated that segments/residues in eIF1, eIF1A, tRNAi and 18S rRNA, which are implicated genetically in TC recruitment, also stimulate the rate of this reaction in a fully reconstituted yeast in vitro translation system. (nih.gov)
  • Th)1 cells.5 Meanwhile, the function and immunity of with genetic predisposition, involving multiple cells, DCs are closely regulated by miRNAs. (cdc.gov)
  • nanos mutations disrupting metal binding at either of these two sites in vitro abolish Nanos translational repression activity in vivo. (embl.de)
  • Their position in the 5' UTR suggests they contain homologous nucleotides with functional equivalency. (news-medical.net)
  • Synthesis of the sgmRNA in coronaviruses requires a discontinuous step guided by a conserved transcription regulatory sequence (TRS) motif, which is located immediately downstream of the leader sequence (TRS-L) and upstream of each structural and accessory protein-encoding gene (TRS-B) [ 5 ]. (biomedcentral.com)
  • Functional analysis of the human TBX20 promoter revealed a 100 bp region that contains strong activating elements. (nih.gov)
  • Within this core promoter region we recognized functional binding sites for TFAP2 transcription factors and identified TFAP2 as repressors of the TBX20 gene in vitro and in vivo. (nih.gov)
  • To increase diagnostic success of Washington University in St. Louis (WUSTL) UDN Phase 2 participants with non-diagnostic clinical and genomic evaluations, we propose performing RNA-Seq transcriptomic analyses, long-read DNA genomic sequencing, and functional studies of noncoding variants in deep intronic or untranslated (5 and 3' UTR) regions. (hhs.gov)
  • PCR-based methods also can detect a much wider range of EV variants, including those within species A, as well as human parechoviruses (HPeVs) that often grow poorly in vitro and which previously have been underdiagnosed by traditional methods. (ox.ac.uk)
  • Methods: 5'-RACE was performed to identify the transcription start site of chicken VNN1. (koreascience.kr)
  • Notch signals are required for in vitro but not in vivo maintenance of human hematopoietic stem cells and delay the appearance of multipotent progenitors. (sunnybrook.ca)
  • Human CD8 T cells generated in vitro from hematopoietic stem cells are functionally mature. (sunnybrook.ca)
  • FDA approval PARP inhibitor Our results indicate that the downregulation of RhoGDI alpha similarly affects the in vitro migration and invasion of ER+ MCF7 and ER- MDA-MB-231 cells. (pdgfrsignals.com)
  • Examples of a novel series of phenanthrolinones are shown to be potent competitive inhibitors of avian prolyl 4-hydroxylase, and of collagen hydroxylation, in embryonic chick tendon cells and human foreskin fibroblasts in vitro and in the oestradiol-stimulated rat uterus in vivo. (embl.de)
  • JASPAR and TFSEARCH were used to analyze the potential transcription factor binding sites in the promoter region of chicken VNN1 and miRanda was used to search miRNA binding sites in 3' untranslated region (3'UTR) of chicken VNN1. (koreascience.kr)
  • Finally, we used siRNAs against the major transcription factors (TFs) predicted for these regions [peroxisome proliferator-activated receptor γ (PPARγ), Krüppel-Like Factor 4 (KLF4), and vitamin D receptor (VDR)] to assess EDN1 expression in cell culture and validate the binding sites. (stanford.edu)
  • Expression of versican 3'-untranslated region modulates endogenous microRNA functions. (sunnybrook.ca)
  • Expression of CD44 3'-untranslated region regulates endogenous microRNA functions in tumorigenesis and angiogenesis. (sunnybrook.ca)
  • There is an increasing evidence that RG4s control gene expression at transcriptional and post- transcriptional levels, although such data is largely based on in vitro studies. (grantome.com)
  • Consistent with the in vitro data, TF expression and TF activity, but not TNFα expression, were increased in Parp14−/− mice in vivo. (imperial.ac.uk)
  • We used a knock-down strategy to manipulate PPARα (or miRNA-181a-5p) expression levels in vitro to further investigate its effect on VNN1 gene transcription. (koreascience.kr)
  • Although more and animals and plants and regulate gene/protein expression more progressions have been made about the through direct complementarity between their 5' region pathogenesis of asthma in recent years, the increasing and the 3' untranslated region of target mRNAs. (cdc.gov)
  • The detection assay includes oligonucleotide primers and dual-labeled hydrolysis (Taqman-style) probes for the in vitro quantifiable detection of HAV. (pagefreezer.com)
  • method II, Express qPCR one-step kit) with primers and probes targeting the EV and HPeV 5'-untranslated regions (5'UTR). (ox.ac.uk)
  • RNA regions containing RG4s significantly overlap with regions containing disease- associated mutations. (grantome.com)
  • In 1987, a region of linkage with AD was reported on the long arm of chromosome 21, which encompassed a region harboring the β- APP gene, a compelling candidate for AD. (touchneurology.com)
  • Albers and colleagues applied high-throughput sequencing in 5 unrelated patients with TAR syndrome and the chromosome 1q21.1 deletion. (medscape.com)
  • [ 5 ] Subsequently, an interstitial microdeletion of chromosome 1q was identified in 30 patients with TAR syndrome. (medscape.com)
  • Data on influenza-like illness (ILI) were obtained from the 13-county Florida Department of Health Region 3 (2010 population 2.2 million) of Florida's Electronic Surveillance System for the Early Notification of Community-based Epidemics ( 3 ). (cdc.gov)
  • Finally, we showed great improvement of the replicon activity and SRIPs production by insertion of a cis-active hammerhead ribozyme (HHRib) before the 5′-untranslated region (UTR). (virosin.org)
  • Platelet interaction and platelet activity in vitro . (leibniz-hki.de)
  • Chen P et al (2012) Molecular determinants of enterovirus 71 viral entry: cleft around GLN-172 on VP1 protein interacts with variable region on scavenge receptor B2. (virosin.org)
  • To this end, the zebrafish ( Danio rerio ) is an established model to study early nervous system development, which combines the complexity of a vertebrate organism with the easy-to-use and high-throughput capabilities of in vitro models ( MacRae and Peterson, 2015 ). (frontiersin.org)
  • Within the C-terminal region are two unusual Cys-Cys-His-Cys (CCHC) motifs that are potential zinc-binding sites. (embl.de)
  • [ 14 ] A region known as the reactive center loop (RCL) protrudes above the core of the serpin molecule and has a sequence of amino acids that is complementary to binding sites in the active sites of the target proteases. (medscape.com)
  • The viral strain has been designated as patients with SARS (4,5). (cdc.gov)
  • The consequences of an A-to-I editing event range from the trivial to the critically important, and the cellular functions of the majority of editing events are still unknown [ 5 ]. (biomedcentral.com)
  • Analysis of nanos mutants reveals that a small, evolutionarily conserved, C-terminal region is essential for Nanos function in vivo, while no other single portion of the Nanos protein is absolutely required. (embl.de)
  • Several polymorphisms were identified in CXCL12 gene including rs1801157 in the 3′-untranslated region, which is characterized by a substitution of a guanine for an adenine. (biomedcentral.com)
  • The kit also includes an In-Fusion Snap Assembly Master Mix for seamless cloning of the target DNA into the pre-linearized vector and a FLuc Control Fragment, consisting of 15 bp 3' and 5' In-Fusion sequences and Fluc CDS (1,683 bp). (takarabio.com)
  • The affinity of hOAT1 toward cidofovir and adefovir ( K m = 46 and 30 μM, respectively) was 5- to 9-fold higher compared with rROAT1 ( K m = 238 and 270 μM, respectively). (aspetjournals.org)
  • Alzheimer's disease (AD) is the most common cause of dementia in the elderly, with a prevalence of 5% after 65 years of age. (touchneurology.com)
  • Specifically, they identified the presence of stem-loops at the 5' end of the RNA genome and other structures involved in frameshifting and replication. (news-medical.net)