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We narrowed to 42 results for: lenti sgrna

Showing: 21 - 40 of 42 results

  2. Viral Vectors 101: Viral Applications

    Type
    Blog Post
    ...shRNA sequence specific to your gene’s mRNA or a sgRNA targeting an exon in your gene of interest, for ...with many applications. These libraries include sgRNAs targeting the entire genome, overexpression of ...is more than one type of virus to choose from (lentivirus, retrovirus, AAV, etc.). When you’ve decided ...

  3. Cre-lox system

    Type
    Collection
    ...60226 AAV:ITR-U6-sgRNA(backbone)-pEFS-Rluc-2A-Cre-WPRE-hGHpA-ITR Cre, luciferase, and sgRNA expression EFS... Zhang 60229 AAV:ITR-U6-sgRNA(backbone)-pCBh-Cre-WPRE-hGHpA-ITR Cre and sgRNA coexpression Cbh AAV Zhang...ITR-U6-sgRNA(backbone)-hSyn-Cre-2A-EGFP-KASH-WPRE-shortPA-ITR Cre, KASH-tagged EGFP, and sgRNA expression...Zhang 60820 pSECC Cre, Cas9 and sgRNA expression EF-1 alpha Lentiviral Jacks 60877 pMAZe Cre and dtomato...12238 pLOX-CW-CRE Cre CMV Lentiviral Trono 12265 pHR-CMV-nlsCRE Cre CMV Lentiviral Trono 12493 p259 pCMV-...coexpression CMV Lentiviral Sadelain 30205 Cre-IRES-PuroR Cre and PuroR coexpression EF-1 alpha Lentiviral Kotton...-cre mCherry and Cre Lentiviral Luikart 66691 FUG-T2A-cre GFP and Cre Lentiviral Luikart 66692 pRubiC-...

  5. What's New in CRISPR - December 2019

    Type
    Blog Post
    ... expression. They replaced the tetraloop of the sgRNA scaffold with a com aptamer which binds an aptamer...Base editors CRISPR toolkit for epitope tagging Lentiviral capsid-based Cas9/gRNA ribonucleoprotein delivery...toolkit   Cas9/gRNA ribonucleoprotein delivery by lentiviral capsid-based bionanoparticle Transient expression... Cas9. Recently, Baisong Lu’s lab developed a lentiviral capsid-based bionanoparticle system for packaging...aptamer binding protein for packaging into a lentivirus-like particle. Read the paper in Nucleic Acids Research...

  6. CRISPR Kinome Libraries Available: Pooled and Individual Plasmid Formats

    Type
    Blog Post
    ...References: 1. Doench, John G., et al. "Optimized sgRNA design to maximize activity and minimize off-target...available in two vector formats: all-in-one (lentiCRISPRv2, expressing both Cas9 and an individual gRNA...gRNA from this single vector) and guide-only (lentiGuide expressing only an gRNA, for use with cells that...at Addgene, you can also order the individual lentiGuide gRNA plasmids targeting the human kinome. These...

  7. CRISPR 101: Off-Target Effects

    Type
    Blog Post
    ...of 5%. Your experiment involves making a large sgRNA library to thousands of genes and targeting and ...sequencing millions of cells. For every individual sgRNA, you recover a minimum of 1,000 targeted cells to...been used - GUIDE-Seq and integrase-defective lentiviral vectors - which rely on insertion of DNA by the...

  8. Tips for a 1st Time CRISPR User (by a 1st Time CRISPR User)

    Type
    Blog Post
    ...is described in Rational design of highly active sgRNAs for CRISPR-Cas9-mediated gene inactivation. Doench...Doench & David Root and are described in Optimized sgRNA design to maximize activity and minimize off-target...expressing Cas9 via transduction with Cas9 lentiviral vector, lentiCas9-Blast (52962-LV, Figure 1). Some cells...Lines with Lentivirus Isolating a Monoclonal Cell Population by Limiting Dilution Lentivirus Production...performing and analyzing a gene edit using Addgene’s lentiviral CRISPR tools. Schematic overview of CRISPR...Cas9 expression in A549 cells transduced with lentiCas9-blast. Anti-Cas9 Western blotting was performed...of A549 cells transduced with Cas9 expressing lentivirus at the indicated multiplicity of infection (MOI...

  9. Sequencing Options for CRISPR Genotyping

    Type
    Blog Post
    ...many software tools predict off-target effects of sgRNAs using computational methods. They identify possible...mismatches based on the sequences of the genome and sgRNA. This is a good starting point for most researchers...CRISPR-Cas9 and TALENs using integrase-defective lentiviral vectors." Nature biotechnology 33.2 (2015): 175... for Your Experiments Get Ready-to-Use CRISPR Lentivirus ...

  10. Viral Vectors 101: Gamma-Retroviral Packaging Systems

    Type
    Blog Post
    ...γ-Retroviral transfer plasmid encoding a transgene, sgRNA, or shRNA of interest: The transgene sequence is... cells a virus can infect. Note: While both lentiviruses and γ-retroviruses use the same gene products... well as the viral LTRs, differ. As a result, lentiviral and γ-retroviral packaging plasmids are not interchangeable...using 293T cells This method is very similar to lentiviral packaging methods. The three components described...How do engineered γ-retroviruses differ from lentiviruses? There are a few important differences between...between engineered γ-retroviruses and lentiviruses. First, these viruses were derived from different genomes...genomes (MoMLV and MSCV for γ-retrovirus; HIV for lentivirus). Additionally, γ-retroviruses can only transduce...

  11. Hot Plasmids - June 2020 - Barcoded CRISPR Library, Sparse Cell Labeling, Calcineurin Reporter, and DNA Staining Dye Alternative

    Type
    Blog Post
    ...deposit please sign up here.  Human whole genome sgRNA iBAR library Article contributed by Susanna Bachle...region. After infecting cells with sgRNAiBAR library containing lentiviral vectors the iBARs can be detected...Initial steps in the CRISPR-pooled screening using sgRNAiBAR. Image from Zhu et al., 2019.  Wensheng Wei's...the gRNAs and their iBAR sequences. Since the sgRNAiBAR library covers every annotated human gene, it ...number of cells and in vivo settings. Find the sgRNAiBAR library!  Zhu et al., Genome Biology, 2019. PubMed...

  12. Pairing CombiGEM and CRISPR for Combinatorial Genetic Screening

    Type
    Blog Post
    ...PMC4151723. 2. Doench, John G., et al. "Optimized sgRNA design to maximize activity and minimize off-target...digestion and ligation steps to build barcoded lentiviral plasmids containing one or more gRNAs. As each..., the ligation products are a diverse pool of lentiviral plasmids. Multiple rounds of digestion and ligation...for CombiGEM-based assembly into the CombiGEM lentiviral vector backbone pAWp12 (Figure 2). Cloning from...from the pAWp28 storage vector to the pAWp12 lentiviral vector should retain the diversity in your gRNA...recessive genetic screening in mammalian cells with a lentiviral CRISPR-guide RNA library." Nature biotechnology...

  13. Luciferase Plasmid Collection

    Type
    Collection
    ...plasmid Attila Karsi 105621 Lenti-luciferase-P2A-Neo Firefly EF1α Lentiviral expression of firefly luciferase...Luciferase Type Promoter Description PI 60226 AAV:ITR-U6-sgRNA(backbone)-pEFS-Rluc-2A-Cre-WPRE-hGHpA-ITR Renilla...fusions using Gateway cloning. Lentival. Mikko Taipale 87078 pLenti6.2-Nanoluc-ccdB NanoLuc® Creation ...fusions using Gateway cloning. Lentiviral. Mikko Taipale 87077 pLenti6.2-Firefly-ccdB Firefly Creation ...transfection Feng Zhang 21471 pLenti PGK V5-LUC Neo (w623-2) Firefly PGK Lentiviral expression of firefly luciferase...luciferase Eric Campeau 21474 pLenti CMV V5-LUC Blast (w567-1) Firefly CMV Lentiviral expression of firefly luciferase...Firefly Ubc Lentiviral expression of firefly luciferase Linzhao Cheng 74444 pLenti.PGK.blast-Renilla_Luciferase...

  14. CRISPR Guide

    Type
    Collection
    ...systems contain two components: a guide RNA ( gRNA or sgRNA ) and a CRISPR-associated endonuclease (Cas enzyme...using a prime editing guide RNA ( pegRNA ). Like sgRNA, pegRNA directs the nickase to the target site by...efficiency PE3 - increased efficiency; an additional sgRNA directs Cas9n to a nearby site, driving mismatch...in nature and is also commonly referred to as an sgRNA. gRNA scaffold sequence The sequence within the ... D. E. (2014). Rational design of highly active sgRNAs for CRISPR-Cas9–mediated gene inactivation. Nature...labeling of genomic loci with dCas9 and engineered sgRNAs using CRISPRainbow. Nature Biotechnology . 34 (...CRISPR libraries What Are Pooled Lentiviral CRISPR Libraries? Pooled lentiviral CRISPR libraries are a heterogeneous...

  15. 27 Hot Plasmids from 2016

    Type
    Blog Post
    ...The library was cloned into the Liu lab’s own pAc-sgRNA-Cas9 insect expression backbone, which expresses...that constitutively express Cas9 with a library of sgRNA expression vectors (containing ten guides against...obstacle. With CRISPR-X, dCas9 is used to recruit an sgRNA with MS2 hairpin binding sites, which then recruit...the authors packaged it into lentivirus and used the resulting lentiviral library to generate a population...Science. 2013. PubMed PMID: 23328393 Wu lab human lentiviral CRISPR library     CRISPR pooled libraries enable...applications. First, GMAP-compatible backbones for lentivirus LV 1-5 and retrovirus RV 2-5 were constructed...Library. This complex second-generation CRISPR lentiviral library targets nearly all human protein-coding...

  16. Lentiviral Vector Guide

    Type
    Guide
    ...Transfer plasmid — contains transgene, sgRNA, or shRNA of interest flanked by LTRs; 8–10 kb packaging capacity... Guides Lentiviral Vector Guide Lentiviral Vector Guide Jump To: Lentiviral Vectors Lentiviral Generations...Read our lentiviral guide to learn about lentiviral components, generations, lentiviral production, and...Browse lentiviral plasmids available at Addgene. Lentivirus Versus Gamma-Retrovirus Lentiviruses and gamma-retroviruses... Things Lentivirus Lentiviral Vector Uses and Overview Overcoming the Challenges of Lentiviral Production...community for decades. Wild-type lentiviruses have been engineered into lentiviral vectors that can be safely...main difference between lentiviruses and gamma-retroviruses is that lentiviruses are capable of infecting...

  17. 22 Hot Plasmid Technologies from 2014

    Type
    Blog Post
    ...vectors and sgRNA libraries expand upon the CRISPR family of plasmids by offering a lentivirus-based mechanism...mechanism for sgRNA delivery and providing a means for large scale functional screens. For more information...., BMC Microbiology 2011 Feb 22;11:38. Lentiviral CRISPR libraries for knockout screening New systems...Pooled Libraries webpage or read our blog post,Lentiviral CRISPR Libraries Enable Genome-Scale, Knockout...CoChR have been deposited to Addgene, including lentiviral and AAV expression vectors for certain variants...purification of Cas9 protein in Rosetta E. coli. New Lentiviral CRISPR activator and repressor plasmids from ...

  18. How to Design Your gRNA for CRISPR Genome Editing

    Type
    Blog Post
    ...Shlyueva D, Müller I, Helin K (2016) Optimizing sgRNA position markedly improves the efficiency of CRISPR... guide (synthetic, in vitro transcription, or lentiviral delivery) can affect the accuracy of a predictive...

  19. CRISPR 101: Multiplex Expression of gRNAs

    Type
    Blog Post
    ...Mutagenesis Using Transgenic Expression of Cas9 and sgRNAs.” Genetics 200(2) (2015): 431-441. PubMed PMID:...Multiple Lentiviral Expression Systems (MuLE) Kit: This kit can be used to create lentiviral vectors expressing...Assembly. Using BsaI, gRNAs are cloned into one of 12 pYLsGRNA plasmids, which contain various promoters and ...CRISPR/Cas9-based genome engineering from a single lentiviral vector.” Nucleic Acids Research 42(19) (2014)...

  20. CRISPR Guide

    Type
    Guide
    ...systems contain two components: a guide RNA ( gRNA or sgRNA ) and a CRISPR-associated endonuclease (Cas enzyme...using a prime editing guide RNA ( pegRNA ). Like sgRNA, pegRNA directs the nickase to the target site by...efficiency PE3 - increased efficiency; an additional sgRNA directs Cas9n to a nearby site, driving mismatch...in nature and is also commonly referred to as an sgRNA. gRNA scaffold sequence The sequence within the ... D. E. (2014). Rational design of highly active sgRNAs for CRISPR-Cas9–mediated gene inactivation. Nature...labeling of genomic loci with dCas9 and engineered sgRNAs using CRISPRainbow. Nature Biotechnology . 34 (...CRISPR libraries What Are Pooled Lentiviral CRISPR Libraries? Pooled lentiviral CRISPR libraries are a heterogeneous...
Showing: 21 - 40 of 42 results