...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...

...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...

... 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...

...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  ...

...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 ...

...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...

... 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...

...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...

...classroom Engineering the Plant Genome Using CRISPR/Cas9 pSiM24: Simplifying Plant Genetic Engineering...

...Therapeutics and is an Addgene depositor. KSQ uses a CRISPR-Cas9 platform to systematically test the function ...

...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...

... Intersectin, Cdc42, Raf1, Mek1, Mek2, Cdk5, and Cas9. Xin has demonstrated the utility of these FLiPs...Photosactivatable Fluorescent Proteins Use Light to Control CRISPR Genome Editing Choose the Brightest Fluorescent...

... 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...

...for endogenous protein fusions created using CRISPR/Cas9. eNLs offer high luminescent signal for imaging...

...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...

...translation in living single cells. However, before CRISPR-Cas9 these techniques were quite tedious and, even...

... libraries Read all CRISPR blog posts Resources on Addgene.org Browse our CRISPR interference plasmids...plasmids Find bacterial CRISPR plasmids Read our CRISPR guide ...about other CRISPR methods for bacterial genome engineering   Learn about human CRISPRko, CRISPRi, and CRISPRa...them. For example, gene knockdown with CRISPR interference (CRISPRi) has been limited to lab-adapted bacteria...disease-associated. Mobile-CRISPRi: a modular approach for CRISPRi transfer Mobile-CRISPRi offers two advantages...knockdown experiments. Mobile-CRISPRi applications Mobile-CRISPRi expands CRISPRi to non-model bacteria including...introduce CRISPRi machinery into diverse bacteria species. Existing protocols can transfer CRISPRi into a...

...inserts (such as CRISPR guides) while a single 72h harvest is best for larger genes such as Cas9. See figure...

...plasmid sharing, the CRISPR community has created hundreds of useful variants of Cas9. By providing access...

...RNA amplification, new toehold switch designs, CRISPR/Cas9-mediated genotyping, as well as our electronic...