Skip to main content
Addgene
Showing: 1 - 20 of 29 results
  1. RNA Interference in Plant Biology: New Tools for an Old Favorite

    Type
    Blog Post
    ...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...
  2. Mammalian RNAi Tools

    Type
    Collection
    ...Kissler 12247 pLVTHM Expresses shRNA under the H1 promoter; shRNA or a H1-shRNA cassette from another vector...causes both EGFP and shRNA to be recombined out of the construct, turning OFF shRNA expression Tyler Jacks... both puromycin and shRNA to be recombined out of the construct, turning OFF shRNA expression Tyler Jacks...browse by constitutive lentiviral shRNA, cre-regulated lentiviral shRNA, and more. Plasmid...for more information on shRNA design and delivery. Constitutive Lentiviral shRNA Lentiviral vectors for...constitutive shRNA expression ID Plasmid Description PI 10878 pLKO.1 ‐ TRC cloning vector shRNA expression...1864 pLKO.1 ‐ scrambled shRNA Negative control vector containing scrambled shRNA David Sabatini 11795 pLL3.7...
  3. Popular Retroviral Vectors and Their Uses in Scientific Research

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

    Type
    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...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

    Type
    Blog Post
    ...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

    Type
    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...glycoprotein (Indiana strain). Use with packaging construct encoding Tat. Reiser 17531 pCD/NL-BH*DDD 2nd Packaging...GP. Good for work in human MSCs. Use packaging construct encoding Tat. Reiser Return to Top Popular Lentiviral...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...
  7. Tetracycline Inducible Expression

    Type
    Collection
    ...pAAV-Ptet-RFP-shR-rtTA AAV; shRNA cloning vector; for evaluation of shRNA efficacy using fluorescence;...for additional constructs TetR On Geley Transactivators (tTA or rtTA) Find a construct that expresses ...Tetracycline-inducible Empty Backbones Find a construct that will allow you to insert and induce your ... lentiviral plasmid for inducible expression of shRNA; neomycin selection; plasmid 21915 has puromycin...vector for inducible expression of transgene or shRNA; see article for detailed information about cloning...Lentiviral vector for inducible expression of a miR-shRNA; selection cassette in format: rtTA3+IRES+Neo; see...On Fraser 44012 pInducer20 Lentiviral vector for shRNA expression; see article for additional tookit plasmids...
  8. Cre-lox system

    Type
    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, Cre expression...switch CMV Mammalian Livet loxP (floxed) Constructs These constructs allow for Cre-regulated gene expression...Depending on the construct, Cre may activate or repress gene expression. Common construct types include:... expression promotes shRNA expression. Gene Switch: These constructs contain two genes of interest, Genes...Blog: FLEx Vectors Cre-containing Plasmids loxP Constructs Cre Reporters & Tools Additional Resources References...recombination principles, scientists have developed constructs to activate/inactivate genes when Cre is present...
  9. Empty Backbones - Choosing Your Perfect Plasmid Backbone

    Type
    Collection
    ... Lentiviral shRNA expression Tet-pLKO-puro - Tet-inducible lentiviral shRNA expression ... shRNA expression Tet-pLKO-puro - Tet-inducible lentiviral shRNA expression ...Retroviral shRNA expression pSico PGK GFP - Conditional shRNA expression through... different genetic element or want to express shRNA. Choose by: Species-specific expression Epitope ...markers Viral expression and packaging Reporters, shRNA expression, transgenics and genome modification...Tet-pLKO-neo - Tet-inducible lentiviral shRNA expression pLenti CMV Neo DEST - ...expression pMKO.1 puro - Retroviral shRNA expression pLenti CMV Puro DEST - Lentiviral...
  10. 28 Hot Plasmid Technologies from 2015

    Type
    Blog Post
    ...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...
  11. 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...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...
  12. Plasmids 101: Gene, enhancer, and promoter trapping

    Type
    Blog Post
    ... 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...
  13. Hot Plasmids - January 2023

    Type
    Blog Post
    ...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...
  14. 22 Hot Plasmid Technologies from 2014

    Type
    Blog Post
    ...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...
  15. Viral Vectors 101: Viral Vector Elements

    Type
    Blog Post
    ...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...
  16. Retrovirus Plasmids

    Type
    Collection
    ...Weinberg 10676 pMKO.1 GFP MoMLV U6-driven plasmid for shRNA expression; also expresses GFP. See pMKO.1 puro ...cytoplasm Hahn 11375 MDH1-PGK-GFP_2.0 MSCV Retroviral construct for miRNA expression Chen 52114 pMSCV-IRES-mCherry...27995 TtRMPVIR CMV/MSV Tet-regulated expression of shRNA; expresses rtTA. See Zuber et al. 2011 for additional...
  17. CRISPR 101: RNA Editing with Cas13

    Type
    Blog Post
    ...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

    Type
    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

    Type
    Blog Post
    ...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 29 results