...similarities and differences to Cas9. DNA targeting Cas enzymes Cas9 – The genome engineering workhorse...CRISPR’ without shortly thereafter hearing ‘Cas9’. The Cas9 nuclease is a genome engineering powerhouse...making Cas9 an adaptable choice for many genome engineering endeavors. For more information on Cas9 variants...same mechanism as traditional spCas9. If you are looking to modify RNA, Cas9 just won’t cut it – literally...We are all oh so familiar with the nuclease Cas9, but how does it stack up to the new Cas enzymes on ...proceed to generate a blunt DNA double strand break. Cas9 has been studied for genome engineering purposes...variants, check out our blog post on choosing a Cas9 enzyme. Fast facts – PAM requirement: NGG and an expanding...

...manipulate the mouse genome using CRISPR/Cas9. Similar to the human genome, the mouse genome is made up of...alternative Cas9, from Streptococcus aureus, is as effective at editing mouse embryos as Cas9 from S. pyogenes...about 125 nt that specifies the target, the Cas9 endonuclease that creates the DNA double-strand break (...guest bloggers, Wenning Qin and Haoyi Wang. CRISPR/Cas9 is revolutionizing the mouse gene-targeting field...robust in performance. In its natural setting, CRISPR-Cas9 is an acquired immune system in bacteria and archaea...model basics To create a mouse model, the gRNA, Cas9, and donor oligonucleotide or plasmid components...AAV can also be used as a vector to deliver CRISPR/Cas9 either through injection into either an oocyte or...

...of this system lies in the ability of the endonuclease Cas9 to create a DNA double stranded break (DSB..., and co-injection of the targeting vector and a Cas9 expression plasmid will insert the desired genetic...create C-terminal tag fusions of endogenous genes in human cells. To use CRISPaint, users need 3 vectors:  ...reactions are then mixed and transfected along with a Cas9 expression vector. They can also be introduced directly...into a Drosophila S2 cell line stably expressing Cas9. This PCR toolkit offers C- and N-terminal tagging... promote homology directed repair after a CRISPR/Cas9 induced break. Learn more about these constructs.... “SapTrap, a Toolkit for High-Throughput CRISPR/Cas9 Gene Modification in Caenorhabditis elegans.” Genetics...

... available with multiple Cas9 variants, including high fidelity Cas9s, Cas9s with various PAM requirements...about Cas9 multiplexing . Browse Plasmids: Multiplex gRNA Vectors Engineering the Cas9 Endonuclease CRISPR...recognition; inhibiting Cas9’s endonuclease domain; or inducing degradation of the Cas9 protein. Most Acr proteins...strand SpCas9-HF1 - disrupt Cas9’s interactions with DNA phosphate backbone HypaCas9 - increase Cas9 proofreading...new tools. Figure 3: Overview of Cas9n Nickase and dead Cas9 The two Cas9 nuclease domains, RuvC and HNH...enhanced specificity by engineering Cas9 into high fidelity Cas9s (hfCas9) . There is currently no defined...application. Cas9 Orthologs While SpCas9 is the most commonly used Cas enzyme, additional Cas9 orthologs ...

...catalytically inactive form of Cas9 known as dCas9 (Jinek et al., 2012). Like Cas9, dCas9 can bind to a specific...editing, numerous discoveries have been made using the Cas9 nuclease in the absence of genomic alterations. ...reported, as well as affinity purification with an anti-Cas9 antibody. These different tag systems and affinity...Using similar affinity purification with an anti-Cas9 antibody, the Li lab found a microRNA (miR483) that...upregulates expression of IGF2, a characteristic of many human malignancies. Additionally, the Tapscott Lab identified...PMCID: PMC3664290.  Zhang, Yiqun, et al. "CRISPR Cas9-guided chromatin immunoprecipitation identifies ..., et al. "A programmable dual-RNA–guided DNA endonuclease in adaptive bacterial immunity." Science337.6096...