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We narrowed to 58 results for: shRNA

Showing: 1 - 20 of 58 results

  1. Plasmids 101: Expressing shRNAs for RNA Interference

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    Blog Post
    ...types, while shRNA and siRNA are synthetic constructs introduced through external methods. shRNA processing...plasmids, shRNAs hijack the endogenous miRNA machinery for processing (Figure 1). The shRNA sequence is...BioRender.com.   Expressing shRNAs using plasmids The design of shRNA plasmids requires three important... specificity. Using your shRNA plasmids One of the most common uses of shRNAs is to generate stable knockout...experiment. For this blog, we’ll be focusing on shRNA, one of the most popular choices for gene silencing... such as: Micro-RNA (miRNA) Short hairpin RNA (shRNA) Short interfering RNA (siRNA) Piwi-interacting ... is transcribed to produce hairpin shRNA structures that are processed by a ribonuclease called Dicer....

  2. Plasmids 101: Control Plasmids

    Type
    Blog Post
    ...variable is the shRNA. Therefore the empty control vector would be Plasmid A sans shRNA, or backbone Y ...Experiment: Knockdown the expression of Gene X using shRNA expressed from a plasmid   Figure 1: Expression ...a single experiment wherein Plasmid A (encoding shRNA against human Gene X in backbone Y) was transfected... this result one could simply conclude that the shRNA didn’t work, as the expression level of Gene X in...plasmid delivered into the cells successfully? Is the shRNA target correct? Is the purified plasmid DNA viable... in the experiment above, Plasmid A contains an shRNA that targets human Gene X and human cells are being...non-targeting control in this experiment could be an shRNA—in the same backbone as Plasmid A—that does not ...

  3. Popular Retroviral Vectors and Their Uses in Scientific Research

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    Blog Post
    ...inducible gene overexpression, knockdown (shRNA, miR-30-shRNA), mutation/editing (CRISPR-Cas9) or deletion...vectors can regulate gene expression by delivering shRNA or miRNA. The ever expanding CRISPR field makes ...in mammalian cells.  Learn more about Mammalian shRNA Tools for RNAi! Cre-Lox technology CAG-GFP-IRES-...empty backbones) here. Cloning and expressing new shRNA sequences pLKO (Plasmid #10878) This plasmid has...Genetic Perturbation Platform, to produce their shRNA library to support functional investigations of ...sequence that can be removed and replaced with an shRNA sequence of your choice. More information and a ...

  4. 15 Years of Addgene: The Top 15 Plasmids

    Type
    Blog Post
    ...lentiviral vector that contains a scrambled shRNA. This scrambled shRNA is used as a negative control for mammalian...expressing plasmid pMD2.G. Find pMDLg/pRPE. scramble shRNA - This plasmid from David Sabatini’s lab is a 3rd...mammalian RNAi experiments.  Find the scrambled shRNA vector. pLKO.1 - TRC cloning vector - This vector...transfer plasmid for cloning and expressing new shRNA sequences. For more information, see the Addgene...particles. It’s capable of inducible expression of shRNA in target cells by the addition of tetracycline/...

  5. Protocol - pLKO.1 – TRC Cloning Vector

    Type
    Protocol
    ...insert. pLKO.1 – scramble shRNA Negative control vector containing scrambled shRNA. psPAX2 Packaging plasmid...Map of pLKO.1 A.3 Related plasmids B. Designing shRNA Oligos for pLKO.1 B.1 Determine the optimal 21-mer... Order oligos compatible with pLKO.1 C. Cloning shRNA oligos into pLKO.1 C.1 Recommended materials C.2...which The RNAi Consortium has built a library of shRNAs directed against 15,000 human and 15,000 mouse ...genes. Addgene is working with the TRC to make this shRNA cloning vector available to the scientific community...lentiviral vector chosen by the TRC for expression of shRNAs. pLKO.1 can be introduced into cells via direct...selection. Figure 1 : Map of pLKO.1 containing an shRNA insert. The original pLKO.1-TRC cloning vector has...

  7. Mammalian RNAi Tools

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    Collection
    ...browse by constitutive lentiviral shRNA, cre-regulated lentiviral shRNA, and more....transcribed, producing the shRNA. Dicer, an RNase III enzyme, binds the shRNA and cleaves it to produce...information on shRNA design and delivery, see resources below. Figure 1: Overview of shRNA-mediated RNA ...BioRender.com. shRNA Plasmids Browse our selection of empty vectors for cloning in shRNAs. Most plasmids... Mammalian RNAi Mammalian shRNA Tools for RNAi Browse Plasmids Resources RNA interference (RNAi) is an... target sequence. The dsRNA can be delivered as shRNA (short hairpin RNA) or siRNA (short interfering ...via transfection or viral delivery of a plasmid. shRNAs are often delivered using viral vectors. Lentiviral...

  8. Lentivirus Plasmids

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    Collection
    ...expression of shRNAs; addition of Cre turns on shRNA expression. See article for more cre-lox shRNA expression...the gene of interest, also can express shRNA (cloning H1-shRNA cassettes into the unique SnaBI site in...Generation Description PI 8453 pLKO.1 puro 3rd U6-driven shRNA empty plasmid with puro resistance. See plasmid ...10878 pLKO.1 – TRC cloning plasmid 3rd U6-driven shRNA empty plasmid; includes a stuffer for easy cloning...cloning Root 14748 pLKO.3G 3rd U6-driven shRNA empty plasmid with EGFP marker. See plasmid 14749 for Thy1.1...21915 Tet-pLKO-puro 3rd inducible expression of shRNA; puromycin selection. See plasmid 21916 for Neo ...Baltimore 12247 pLVTHM 2nd EF-1a-driven GFP and shRNA under the control of a tet-responsive H1 promoter...

  9. RNA Interference in Plant Biology: New Tools for an Old Favorite

    Type
    Blog Post
    ... address these shortcomings, both short hpRNAs (shRNAs) and artificial micro RNAs (amiRNAs) are frequently...precursors of microRNAs found in the genome, whereas shRNAs are exogenously derived. Importantly, instead of...single ~21-nt interfering RNA is produced using shRNA/amiRNA. Although sh- and amiRNAs are likely more...Arabidopsis species, respectively. Short hairpin RNA (shRNA) is conceptually similar to the microRNA precursor...research community. Fortunately, many tools for shRNA in mammals are available at Addgene. Identifying...

  10. Viral Vectors 101: Viral Applications

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    Blog Post
    ...several options to allow you to do exactly that: shRNAs are short RNA sequences which target complementary...sequence for degradation. Viral introduction of shRNA plasmids can induce gene-specific silencing temporarily...the viral vector for your gene of interest – an shRNA sequence specific to your gene’s mRNA or a sgRNA...

  11. Empty Backbones - Choosing Your Perfect Plasmid Backbone

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    Collection
    ...basal activity shRNA/RNAi Gene silencing Browse RNAi empty vectors for cloning in shRNAs, including plasmids...Tet-inducible lentiviral shRNA expression pLKO.1 - TRC cloning vector - Lentiviral shRNA expression pLJM1-EGFP...a different genetic element, or want to express shRNA. Choose by: Species-Specific Expression Epitope ...delivery, genome modification, reporter assays, shRNA expression, transgenics, and more. See the next ...retroviral delivery of one sgRNA pMKO.1 GFP - Retroviral shRNA expression Retroviral backbones made by Addgene ...uses such as genome modification, reporter assays, shRNA expression, transgenics, and more. Element Details...Tet-pLKO-puro and Tet-pLKO-neo - Tet-inducible lentiviral shRNA expression pnCasPA-BEC - A cytidine deaminase-mediated...

  12. How to Deposit Your Plasmids with Addgene

    Type
    Blog Post
    ...each insert.  Encodes gRNA/shRNA A plasmid with one or more gRNA or shRNA sequences cloned into it. gRNAs...location, while shRNAs are used to knockdown expression of mRNAs with complementarity to the shRNA sequence....contained within your insert (or targeted by a gRNA/shRNA) and select the species that the gene or insert ...

  13. Plasmids 101: The Promoter Region – Let's Go!

    Type
    Blog Post
    ...an RNAP II promoter, whereas small RNAs (such as shRNA) are transcribed from the RNAP III promoters. This...expression in plants. H1 small RNA expression shRNA From the human polymerase III RNA promoter Constitutive...expression in neuronal cells. U6 small RNA expression shRNA From the human U6 small nuclear promoter Constitutive...

  17. Tetracycline Inducible Expression

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    Collection
    ... pAAV-Ptet-RFP-shR-rtTA AAV Tet-On shRNA vector. To evaluate shRNA efficacy by fluorescence, use Plasmid...Promoters Blog: Repressible Promoters Mammalian shRNA Plasmids Collection Cre-lox Plasmids Collection ...Lentiviral Tet-On plasmid for inducible expression of shRNA with puromycin selection. See Plasmid #21916 for...plasmid for easier cloning and design of inducible shRNA expression, with puromycin selection. See Plasmid... al., 2011 ) for additional tookit plasmids for shRNA or gene expression. rtTA-Advanced H1-2O2 Stephen...vector for inducible expression of transgene and/or shRNA. See article ( Szulc et al., 2006 ) for Tet-Off ...

  18. Cre-lox system

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    Collection
    ...Cre-dependent shRNA Expression: Cre-lox can be used to turn shRNA constructs on or off. In floxed-shRNA constructs... excise the shRNA to return gene expression to physiological levels. In lox-STOP-lox shRNA constructs,...Mammalian Sofroniew 25997 LV-Cre pLKO.1 Cre and shRNA coexpression CMV Lentiviral Fuchs 26646 pCAG-Cre-IRES2... expression and tetracyclin-dependent transgene/shRNA expression ApoE.HCR.hAAT Mammalian Ehmer 85578 pTC-ApoE-Tet... expression and tetracyclin-dependent transgene/shRNA expression ApoE.HCR.hAAT Mammalian Ehmer 85797 pSkipFlox...constructs, Cre expression promotes shRNA expression. Gene Switch: These constructs contain two genes of interest... Expression System PI 11578 pSico Cre activates shRNA expression, removal of EGFP; See also similar plasmids...

  19. CRISPR 101: Targeting RNA with Cas13a (C2c2)

    Type
    Blog Post
    ... size distribution. When comparing LwaCas13a to shRNA, they found that LwaCas13a displays similar knockdown...a distinct contrast to the hundreds of observed shRNA off-targets. They subsequently used catalytically...

  20. Retrovirus Plasmids

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    Collection
    ...Weinberg 10676 pMKO.1 GFP MoMLV U6-driven plasmid for shRNA expression; also expresses GFP. See pMKO.1 puro ...27995 TtRMPVIR CMV/MSV Tet-regulated expression of shRNA; expresses rtTA. See Zuber et al. 2011 for additional...
Showing: 1 - 20 of 58 results