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We narrowed to 32 results for: shRNA construct

Showing: 1 - 20 of 32 results
  1. Plasmids 101: Expressing shRNAs for RNA Interference

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    ...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...(2006). Design and cloning strategies for constructing shRNA expression vectors. BMC Biotechnology, 6(...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 ...
  2. RNA Interference in Plant Biology: New Tools for an Old Favorite

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    ...research and many different constructs are available to the community. Each construct has unique features, such...RNA (hpRNA). This method is facilitated by DNA constructs that enable insertion of ~400 bp complementary...sequence (discussed below). Examples of these constructs include pGAPi, which allows easy creation of ... 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...
  3. Popular Retroviral Vectors and Their Uses in Scientific Research

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    ...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 ...This modular and flexible system can be used to construct complex lentiviruses for modification of mammalian...
  4. Viral Vectors 101: Viral Applications

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    ...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...selectable marker. This vector can express your construct in target cells transiently or constitutively ...the viral vector for your gene of interest – an shRNA sequence specific to your gene’s mRNA or a sgRNA...Some customization is required, but many viral constructs have already been built and are available on ... embryonic stem cells. Viral integration of a construct is also reliably higher than traditional transfection...
  5. The 10 Most Distributed Plasmid Technologies in Addgene's First 10 Years

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    ...item of 2005. shRNA - In 2006, we saw an increase in popularity of empty backbones for shRNA expression,...plasmids you'll note that those are lentiviral constructs. In 2007 we continued to see requests grow for...beta-catenin reporter M50 Super 8x TOPFlash, scramble shRNA for use as a negative control, and empty backbones...
  6. Lentivirus Plasmids

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    ...(Cre-lox), stable expression of shRNAs; addition of Cre turns on shRNA expression. See Ventura et al. ...the gene of interest. Can also express shRNA (cloning H1-shRNA cassettes into the unique SnaBI site in...Transfer plasmid — contains transgene, sgRNA, or shRNA of interest flanked by LTRs Packaging plasmid — ...Generation Description PI 8453 pLKO.1 puro 3rd U6-driven shRNA empty plasmid with puromycin selection. See plasmid...10878 pLKO.1 – TRC cloning plasmid 3rd U6-driven shRNA empty plasmid. Includes a stuffer for easy cloning...cloning. David Root 14748 pLKO.3G 3rd U6-driven shRNA empty plasmid with EGFP marker. See plasmid 14749 for...21915 Tet-pLKO-puro 3rd Inducible expression of shRNA with puromycin selection. See plasmid 21916 for ...
  7. Cre-Lox and Other Site-Specific Recombinases

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    ...deleted. 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, Cre expression... expression promotes shRNA expression. Gene Switch: These constructs contain two genes of interest, Genes...regulate your gene of interest. Depending on the construct, gene expression may be activated or repressed...
  8. 28 Hot Plasmid Technologies from 2015

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    ...sgRNA (U6) expression, shRNA/shRNA-miR30 constitutive (H1, U6, 7SK) or inducible shRNA-miR30 (CMV-TO) expression...expression, or a promoterless construct. Pre-constructed Entry vectors containing Cas9, EGFP, mCherry, iRFP...constitutive or inducible, gene knockdown with shRNA or shRNA-miR30, (2) tamoxifen-inducible CreERT2 gene...of inducible promoter constructs. Up to 34 building blocks were used to construct 20kb long inserts. The...att sites are supplied as individual reporter constructs or as the fully assembled pMemoryArray.   Yang...Biotechnol Lett. 2015. 37:383-389. Toolkit for constructing fusion proteins with customizable N- and C-terminal... to 5 different elements. Lentiviral vectors constructed with the MuLE toolbox have been used for a variety...
  9. Plasmids 101: The Promoter Region – Let's Go!

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    ...an RNAP II promoter, whereas small RNAs (such as shRNA) are transcribed from the RNAP III promoters. This...commonly employed in lentiviral and retroviral constructs and we will discuss these in a future post on...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...
  10. Plasmids 101: Gene, enhancer, and promoter trapping

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    ... rely on the random integration of a reporter construct in close proximity to an endogenous gene. Each...detail below), but fundamentally the reporter construct monitors the expression of a nearby gene. The ...an uncharacterized gene. The actual reporter construct can be something phenotypic (such as a fluorescent...the expression of the gene (by CRISPR knockout, shRNA depletion, etc.) and ask what the phenotype is without...monitor. Gene traps Gene traps are promotor-less constructs that contain splice acceptors. They require insertion...type of trap, there are many ways to insert your construct to generate a collection of monitored genes and...
  11. Retrovirus Plasmids

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    ...Transfer plasmid — contains transgene, sgRNA, or shRNA of interest flanked by LTRs Packaging plasmid — ...Weinberg 10676 pMKO.1 GFP MoMLV U6-driven plasmid for shRNA expression with GFP expression. See plasmid 8452...William Hahn 11375 MDH1-PGK-GFP_2.0 MSCV Retroviral construct for miRNA expression. Chang-Zheng Chen 52114 pMSCV-IRES-mCherry...27995 TtRMPVIR CMV/MSV Tet-regulated expression of shRNA. Expresses rtTA. See Zuber et al. (2011) for additional...
  12. Empty Backbones - Choosing Your Perfect Plasmid Backbone

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    ...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...
  13. Tetracycline Inducible Expression

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    ... pAAV-Ptet-RFP-shR-rtTA AAV Tet-On shRNA vector. To evaluate shRNA efficacy by fluorescence, use Plasmid...Tetracycline-inducible Backbones Find backbones or constructs to use for cloning to control expression of your...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 ...Return to Top Transactivators (tTA or rtTA) Find a construct that expresses the transactivator for your tetracycline...
  14. Hot Plasmids - January 2023

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    ...targeting system Heat-inducible lentiviral Car constructs Anti-GFP [N86/8R] ready-to-use antibody Hyperfolder...endogenous RNA imaging tool.   Fig. 1: Csm, shRNA, and Cas13 were compared head-to-head for gene expression... 36690762.   Heat-inducible lentiviral CAR constructs by: Brian O'Neill  A clever system for reducing...system is a set of heat-inducible lentiviral CAR constructs, the most basic of which is depicted here: ...days - after just 15 minutes of FUS.  Similar constructs to the one above can be made for expressing your...
  15. 22 Hot Plasmid Technologies from 2014

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    ...assembled into a final transgenesis construct. Plasmids constructed using Golden GATEway cloning have been...complexity of assembling recombination of transgenesis constructs? A new cloning system combining the advantages...simplifies the cloning process for complex DNA constructs, particular for those involving recombination...Multisite Gateway cloning to generate the final construct. Up to eight entry vectors can be used for ...methods, particularly for complex transgenesis constructs. Kirchmaier et al., PLOS One 2013 Oct 7;8(10...The Janovjak lab has deposited their 3 Opto-RTK constructs – Opto-mFGFR1, Opto-hEGFR, and Opto-hRET – as...as well as various LOV-domain-mVenus constructs used in the study for those who would like to extend this...
  16. Viral Vectors 101: Viral Vector Elements

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    ...typically an expression construct, to cells/organisms. This is often cDNA, shRNA, or something similar,...), it’s still used to deliver gene expression constructs. Sandwiched between two inverted terminal repeats...just like AAV, which encompass the expression construct and promoter that you wish to deliver to your ...plasmids in our collection. Want to make a vector construct on your own? No problem, we are here to help with...
  17. CRISPR 101: RNA Editing with Cas13

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    ...shrink the construct. They identified ADAR2DD(delta984-1090), which decreases the REPAIRv1 construct size from... A->G editing, they employed hyperactive ADAR constructs, like ADAR2DD(E488Q). They also adjusted the ...Although REPAIRv1 knockdown is more precise than shRNA knockdown, it still displays substantial off-target...binding to decrease off-targets. Of the tested constructs, ADAR2DD(E488Q/T375G) (REPAIRv2) had the highest...dRanCas13b truncations that would allow packaging the construct for viral delivery. They found C-terminal truncations...
  18. Your Lentiviral Plasmid FAQs Answered

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    Blog Post
    ...of viral constructs is a useful tool as it provides a means to quickly check that a construct is functional...variety of applications including cDNA expression, shRNA-mediated knockdown, Tet and Cre-regulated expression...making a stable cell line. When expressing a viral construct transiently care needs to be taken to ensure that...
  19. Viral Vectors 101: Gamma-Retroviral Packaging Systems

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    ...transfer plasmid encoding a transgene, sgRNA, or shRNA of interest: The transgene sequence is flanked by...wildtype LTR promoter is no longer present in these constructs. Why do many γ-retroviral plasmids have similar...
Showing: 1 - 20 of 32 results