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We narrowed to 243 results for: crispr cas9

Showing: 201 - 220 of 243 results

  1. Hot Plasmids February 2024

    Type
    Blog Post
    Published
    Feb. 20, 2024, 2:45 p.m.
    ...cells By Susanna Stroik Even with the advances in CRISPR technology over the past decade, primary cells ...peptide” that facilitates endosomal escape. Combining Cas9-PAGE with lentiviral sgRNA expression yielded editing...lines and primary cells. Although cell entry of a Cas9 ribonucleoprotein (RNP) was limited (likely due ...multiplexed guides. Figure 1: A) Cell-penetrating Cas9, fused to HIV TAT, Myc and SV40 Nuclear Localization...Localization Signals, and GFP. B) Workflow to assess Cas9-PAGE editing with viral sgRNA targeting mCherry reporter...

  2. Pooled Library Amplifications

    Type
    Blog Post
    Published
    May 30, 2024, 1:30 p.m.
    ...Protocol (DOCX, 17 KB)   For CRISPR libraries, Genome-scale CRISPR-Cas9 knockout and transcriptional ...Amplification Type Link to Protocol CRISPR Knockout Human CRISPR Knockout Pooled Library (Brunello) ...Amplification Protocol (PDF, 88.2 KB) CRISPR Knockout Human CRISPR Knockout Pooled Library (GeCKO v2) ... Protocol (DOCX, 15 KB) CRISPR Activation Human Genome-wide CRISPRa-v2 Libraries 104,540–209,080...Re-Amplification Protocol (PDF, 71 KB) CRISPR Activation Human Subpooled CRISPRa-v2 Libraries 12,285–20,145 ...ng Solid Joung et al. Nat Protoc. 2017 CRISPR Knockout Human Epigenetic Knockout Library ...on the Addgene blog Genome-Wide Screening Using CRISPR Preparing Pooled Libraries NGS Quality Control ...

  3. Hot Plasmids - May 2022

    Type
    Blog Post
    Published
    May 26, 2022, 1:15 p.m.
    ...efficiency of up to 20%, comparable to commonly used CRISPR/Cas9 methods for large inserts. However, this system...in comparison to the 62-90% error rate seen with Cas9 editors. The lab believes that this system could...inactive dCas9 and microbial SSAPs (dCas9-SSAP) which insert genes in a cleavage free manner. The dCas9-SSAP...cleavage-free dCas9-based gene editor using microbial SSAPs. Image from Wang, et al, 2022.  Find dCas9-SSAP...Cre-mediated AAV Targeted Evolution method (M-CREATE) dCas9-SSAP with reduced error rate for long sequence knock-ins... https://doi.org/10.1038/s41593-021-00969-4.   dCas9-SSAP with reduced error rate for long sequence knock-ins...Wilson Le Cong’s lab has developed a method using dCas9-SSAP to knock-in long sequences with reduced risks...

  4. Lentiviral Vector Uses and Overview

    Type
    Blog Post
    Published
    May 19, 2016, 2:30 p.m.
    ...Screening Read about Genome-Wide Screening Using CRISPR/Cas9 Additional Resources on Addgene.org Check out...Lentiviral vectors are the vector of choice for many CRISPR applications, and they’ve also had success in clinical...transgene-expressing or knockout cell lines. Many CRISPR plasmids are designed for lentiviral use, including... Improved vectors and genome-wide libraries for CRISPR screening. Sanjana NE, Shalem O, Zhang F. Nat Methods...Lentiviral Plasmid FAQs Answered Learn How Lentiviral CRISPR Libraries Enable Genome-Scale, Knockout Screening...including Addgene’s top requested plasmid in 2015: lentiCRISPR v2, used both for targeted modification and genome-wide...

  5. Cultivating Community Science at BosLab

    Type
    Blog Post
    Published
    Nov. 2, 2017, 1:35 p.m.
    ...our journal club discussion on Kevin Esvelt’s CRISPR/Cas9 gene drive paper was very popular. People kept...Blog Experimenting in an Open Source Lab: from CRISPR to Cats Stem Cell Models for Disease & Open Science...

  6. Cloning Mammalian Cells with the Agarose Method

    Type
    Blog Post
    Published
    Sept. 7, 2017, 12:17 p.m.
    ...48-well plate for further analysis. To assess CRISPR/Cas9 knockout efficacy, we have used both Western ...Addgene Blog Tips for a 1st Time CRISPR User (by a 1st Time CRISPR User) The Challenges of Cell Culture...; more recently we have also been using Addgene CRISPR vectors to generate cell lines with knockouts of...

  7. How Addgene Creates Content

    Type
    Blog Post
    Published
    Nov. 17, 2022, 2:15 p.m.
    ...knowledge changes and require updating or retiring. CRISPR/Cas9, for example, has gone through rapid change in...Resources on the Addgene Blog Science careers Plasmids CRISPR Viral vectors Fluorescent proteins  ...

  8. AAV Q&A with Tim Miles

    Type
    Blog Post
    Published
    May 17, 2022, 1:15 p.m.
    ... primer for beginners Adenoviral delivery of CRISPR/Cas9 Tips for a first time AAV user Resources on .... The continued development of more and smaller CRISPR tools is an exciting area for future AAV application...

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

    Type
    Blog Post
    Published
    Oct. 27, 2020, 1:15 p.m.
    ...RNAs.   RNAi vs. CRISPR: Complementary techniques It is fair to say that CRISPR has revolutionized ...A, Bassik MC (2016) Systematic comparison of CRISPR/Cas9 and RNAi screens for essential genes. Nat Biotechnol...unprecedented ways. With the meteoric rise and diversity of CRISPR technologies, one might incorrectly relegate RNAi...straightforwardness of RNAi is unmatched—unlike CRISPR experiments that require exogenous protein, successful...only requires the presence of dsRNA. In plants, CRISPR technologies are predominantly used to disrupt ...function at the level of DNA, thereby limiting CRISPR’s effectiveness at high copy numbers and genomic...prevalent assumption into question. Specifically, CRISPR-generated mutants believed to be knockouts can ...

  10. Antibodies 101: Validation

    Type
    Blog Post
    Published
    March 24, 2022, 1:15 p.m.
    ...RNAi to knock-down expression of your target or CRISPR/Cas9 to create mutations that essentially eliminate...gene for your protein has been mutated using CRISPR/Cas9. You check the mutation and it causes a large...

  11. Plasmids 101: In-house NGS Plasmid Quality Control Now at Addgene

    Type
    Blog Post
    Published
    April 6, 2021, 12:30 p.m.
    ... MD, Sanjana NE, Zhang F (2017) Genome-scale CRISPR-Cas9 knockout and transcriptional activation screening... of the plasmid in the pool - ex. the gRNA in a CRISPR library. In general, our QC scientists work with...

  12. Hot Plasmids June 2018 - Reverse transcriptase, nanobody, and protein-DNA interaction tools

    Type
    Blog Post
    Published
    June 27, 2018, 1:16 p.m.
    ...lentiviral plasmids, catalytically inactive Cas9 (dSpyCas9) fused to APEX2 is expressed from a Tet/Shield1...Sam Myers, a Scientist in the group, developed a CRISPR-based approach to detect proteins that associate...genomic loci. He made use of catalytically inactive dCas9 by fusing it to an engineered peroxidase, (APEX)...are enriched in the 20 nm radius surrounding the dSpyCas9-APEX2 fusion.  This Sontheimer lab used this technology... technology, which they call C-BERST (dCas9–APEX2 biotinylation at genomic elements by restricted spatial...

  14. Meet the New Addgene Board Members!

    Type
    Blog Post
    Published
    April 28, 2020, 1:14 p.m.
    ...Therapeutics and is an Addgene depositor. KSQ uses a CRISPR-Cas9 platform to systematically test the function ...

  15. Four Ways to Package Transgenes That Exceed the Size Limit of Adeno-associated Virus

    Type
    Blog Post
    Published
    Sept. 1, 2020, 1:15 p.m.
    ...sequential homology directed repairs (HDR) with CRISPR/Cas9. Donor A contains: 400bp homology arms to the...increasing the size of the AAV donor template for CRISPR-mediated homology directed repairs (HDR). Split...single step and serve as donors for two consecutive CRISPR-mediated HDR events which results in the integration....org/10.1172/jci34316 Bak RO, Porteus MH (2017) CRISPR-Mediated Integration of Large Gene Cassettes Using...

  17. Improved Plasmid Maps Powered by SnapGene

    Type
    Blog Post
    Published
    June 29, 2017, 1:06 p.m.
    ... EGFP and mEGFP. Many of these features, such as Cas9, weren’t detected by our in-house software. In fact...are commonly used for Golden Gate cloning or for CRISPR gRNA cloning. With this more functional display...

  18. Luminescent Imaging with Nano-lanterns

    Type
    Blog Post
    Published
    May 25, 2017, 2:30 p.m.
    ...for endogenous protein fusions created using CRISPR/Cas9. eNLs offer high luminescent signal for imaging...

  19. Fluorescent Proteins 101: When GFP lets you down

    Type
    Blog Post
    Published
    Aug. 23, 2018, 12:05 p.m.
    ...nuclease-deficient Cas9 can bind loci in the genome that are defined by a gRNA. Since Cas9 is a protein, a...genomic loci in living human cells by an optimized CRISPR/Cas system." Cell 155.7 (2013): 1479-1491. PubMed...

  20. Visualizing Protein Turnover In Situ

    Type
    Blog Post
    Published
    Jan. 16, 2018, 3:20 p.m.
    ...translation in living single cells. However, before CRISPR-Cas9 these techniques were quite tedious and, even...
Showing: 201 - 220 of 243 results