...Addgene Blog Easi-CRISPR: Generating Knock-In and Conditional Mouse Models CRISPR 101: Cas9 Nickase Design...cells Resources on Addgene.org Browse all CRISPR plasmids CRISPR plasmids for protein tagging Find validated...degradation of FKBP12F36V-BRD4 using CRISPR/Cas9-mediated knock-in, and to evaluate in vivo degradation....ENL-FKBP12F36V and endogenous ENL was inactivated using CRISPR/Cas9 gene-editing. Featured molecules: dTAG-7 and...-MELK and endogenous MELK was inactivated using CRISPR/Cas9 gene-editing. Featured molecules: dTAG-7, ...were engineered to express YY1-FKBP12F36V using CRISPR/Cas9-mediated knock-in. Featured molecule: dTAG...available from Addgene. These plasmids enable lentiviral expression of N- or C-terminally FKBP12F36V-tagged...

...discovery. CRISPR is a great example (the top requested plasmids from 2015 and 2016 were CRISPR plasmids...ubiquitin ligase, cellular senescence, autophagy, CRISPR, and so much more (find the full, searchable list... of 2017 is the lentivirus envelope plasmid pMD2.G from Didier Trono’s lab! Lentivirus envelope vector... producing our ready-to-use lentivirus preps. You can find our lentivirus production protocol on our website...titering your lentivirus in another blog post. If you need a quick primer on how lentiviruses work, check...plasmids), but so are lentiviral vectors, many of which are used to deliver Cas9 and other genes to mammalian...promoter. Envelope proteins are a key component of lentiviruses. These proteins coat the virus and help determine...

...check out our updated CRISPR-Cas resources at www.addgene.org/CRISPR/ to browse CRISPR plasmids, watch informational...For more information on these new CRISPR screening tools, see our CRISPR/Cas Plasmids: Pooled Libraries ...for background information on CRISPR technology? See our improved CRISPR-Cas Guide. Wang et al., Science...Tet-inducible CRISPR activators and repressor plasmids. (Kearns et al., Development. 2014..) CRISPRs for Xenopus.... 2014..) Our CRISPR-Cas collection of plasmids updates frequently, so visit our CRISPR pages often to...BMC Microbiology 2011 Feb 22;11:38. Lentiviral CRISPR libraries for knockout screening New systems...libraries expand upon the CRISPR family of plasmids by offering a lentivirus-based mechanism for sgRNA...

...design your CRISPR gRNA CRISPR 101: Ribonucleoprotein (RNP) Delivery Components of CRISPR/Cas9 ...View Addgene's CRISPR resources Request ready-to-use viral preps of select CRISPR lentiviral plasmids Additional...sequence for CRISPR-Cas9 targeting is led by a ~20 bp gRNA which is part of the larger CRISPR RNA (crRNA...surprise then that stability of the RNA used in CRISPR-Cas9 experiments – the “gRNA” – suffers from the...modifications listed above were developed before CRISPR-Cas9, for use in applications like siRNA and antisense...to find pre-tested gRNA sequences for your next CRISPR experiment.    Chemical RNA modifications... On-off switches for gRNAs A major drawback of CRISPR-Cas9 technology is that most expression systems...

...protocols, and more. Selecting crispr in Guides will return links including our CRISPR guide.  For a closer look...for “CRISPR” plasmids keeps more than half of these. Instead we can navigate first to the CRISPR Resources... search field front and center. Let’s type in “crispr” and see what comes up. Before even clicking the...Type” you can filter by experimental purpose (e.g. CRISPR) Beyond search What can you do if you don’t have...Resources page: The page links to guides, our CRISPR eBook, plasmid kits, and curated pages categorized ...parentheses.  In this example I wanted to find a lentiviral plasmid to express ubiquitin under control of...300+ plasmids. Under ‘Vector Types’ I selected ‘Lentiviral’ which eliminates most of the plasmids designed...

...Addgene.org Check Out Our CRISPR Guide Pages Find Ready-to-Use CRISPR Lentivirus Learn about Other Members...had on the podcast, has worked extensively with CRISPR, and recently became a member of the Addgene advisory...getting an MD PhD, the inside scoop on developing new CRISPR tools, and the potential uses of gene drives. ...in DNA repair informed his decision to work with CRISPR during his postdoc 12:01 - 13:21 : The Wyss Institute...07 : Alex's projects at the Wyss Institute - New CRISPR tools and the importance of moving on from "failed... work and what work he's planning on doing with CRISPR-based gene drives focusing on safety and the ability... crispr...

...our collection. CRISPR/Cas9 - At Addgene, 2013 will be known as the "Year of the CRISPR." The newest genome...many different purposes that we had to create a CRISPR page to keep up with all the new plasmids. So far... far, we have distributed over 10,000 CRISPR/Cas9 plasmids to more than 3,000 labs around the world. ... those are lentiviral constructs. In 2007 we continued to see requests grow for lentiviral packaging and... pLKO.1 puro and pLKO.1 – TRC cloning vector. Lentiviral Plasmids -  If you're familiar with the pLKO ...-puro for retroviral expression and pLVTHM, a lentiviral plasmid for mammalian gene silencing. TALENs ...

...the Addgene blog CRISPR101: Editing the Epigenome  Mapping the 4D Nucleome with CRISPR/Cas9 Resources ...introduce you to a simpler, yet more effective, CRISPR-based epigenetic approach (spoiler alert: it’s ...dioxygenase  When I started to work on building a CRISPR/dCas9-based tool for DNA demethylation in 2015,...al. 2018, Kaas et al. 2013). Figure 2: CRISPR TET-based epigenetic editing. In reality, editing...Moshe Szyf at McGill University, working to develop CRISPR-based tools for epigenetic editing.   References...Resources at Addgene.org Read our CRISPR Guide to Epigenetics References Charlton J, Jung EJ, Mattei AL et al...targeted steric blockage of DNA methyltransferase with CRISPR/dCas9. Nat Commun 12, 5711. https://doi.org/10.1038...

...new Viral Service, we’re distributing lentivirus (with many CRISPR tools included among the preps that ...are used to produce CRISPR tools, we’re starting the Viral Service with lentiviral preps for the delivery.... Of course, lentiviruses are used in experiments other than just those involving CRISPR and so we also... activators, Cas9 repressors, select gRNAs, and CRISPR pooled libraries. Whether you’d like to knockout... Service Suggestion Form. Lentivirus As some of the most popular lentiviral vectors in the repository ...collection. We’ll provide a brief rundown of what lentiviral and AAV preps are available below, but you can...also have lentivirus for delivering control shRNAs, GFP, and rtTA from the Tet-on system. The first two...

... Addgene Blog CRISPR libraries Genome-wide screens with CRISPR Lentiviral CRISPR libraries ...with small molecules/drugs. With the advent of CRISPR, genome-wide knockout screens are possible with...While the entire genome can be knocked out with CRISPR, the entirety of the small molecule realm is typically...downstream manipulations as a forward or reverse screen (CRISPR, drug, etc.), just with an added baseline component...component. Alternative screening techniques While CRISPR and drugs dominate the screening scene, there are... worth considering. RNAi screens are similar to CRISPR, but instead of knocking out the genes of interest... be lethal to cells and thus not assayable with CRISPR. Another method, popularized in yeast, are temperature...