Sigma Aldrich is proud to offer its newest line of genome editing tools, Sigma CRISPRs, to the global research community. As the first company to commercially offer targeted genome editing technology nearly ten years ago, no one has more expertise in this field than Sigma Aldrich. This experience is especially important when it comes to crafting genome editing tools that possess the critical requirements of having specific targeting and robust cutting activity. We provide both in our new Sigma CRISPR product line, and now we put our skill into your hands with a quick and simple web-based design platform. Sigma CRISPR products can be ordered directly through the link below or browse the CRISPR content on this page to learn more about the technology. Sign up to receive the latest updates on CRISPR technology such as new journal articles, protocols, and product launches. On this page: All-in-one, ready-to-use Cas9 and guide RNA (gRNA) expression plasmids CRISPR paired nickases Purified RNA-only guide RNA CRISPR Lentivirus CRISPR positive and negative controls What is CRISPR/Cas? Paired Cas9 Nickases – For optimal Cas9-D10A paired nickase functionality, guide RNAs should be designed in a 5’-to-5’ orientation with PAM spacing between 30 and 150 bp. Sigma Lentiviral CRISPR: (A) Vector map of the all-in-one lenti CRISPR vector. (B) GFP signal post-transduction of HEK293 cells with particles made via pLV-U6g-EPCG. (C) HeLa cells were transduced with lentivirus (pLV-U6g-EPCG) followed by selection on puromycin and 6TG treatment to enrich for HPRT1 knockouts. Insertions and deletions were detected at the gRNA target site via mismatch assay (CEL-I). CEL I image of CRISPR01 positive control (lane labeled EMX1s4+Cas9) and CRISPR02 positive control (lane labeled EMX1s4, EMX1as4+D10A) CRISPR stands for Clustered Regularly Interspaced Short Palindromic Repeats. The discovery of the type II prokaryotic CRISPR “immune system” has allowed for the development for an RNA-guided genome editing tool that is simple, easy and quick to implement. The CRISPR pathway was discovered in bacteria, where it functions much like an immune system against invading viruses and plasmid DNA. Short DNA sequences (spacers) from invading viruses are incorporated at CRISPR loci within the bacterial genome and serve as “memory” of previous infection. Re-infection triggers the complementary mature CRISPR RNA (crRNA) to find a matching sequence – which provides the CRISPR-associated (Cas) nuclease the specificity to form a double-strand break at specific “foreign” DNA sequences. The system is simple, as it only requires a Cas nuclease and a gRNA against the target sequence to function as a site-specific nuclease. Also, despite the bacterial evolutionary origins of the system, data demonstrates high levels of cutting activity in mammalian cells (Cong et al., 2013, Mali et al., 2013), particularly at numerous simultaneous targets, (Wang et al., 2013). In addition, the requirement for an NGG sequence makes target design simple and straightforward in genomic regions where off-targeting is not an issue. Finally, CRISPR provides researchers a fast and cost-effective genome editing tool to use for modifying the genomes of various organisms. Copyright © 2016 Sigma-Aldrich Co. LLC. All Rights Reserved. Reproduction of any materials from the site is strictly forbidden without permission. Sigma-Aldrich Products are sold exclusively through Sigma-Aldrich, Inc. FLUKA, TraceSELECT, TraceSELECT ULTRA, PERDROGEN, CHROMASOLV, FIXANAL, HYDRANAL, Riedel-de Haën, IDRANAL, SPECTRANAL, and VOLPAC are trademarks of Honeywell Specialty Chemicals Seelze GmbH. Site Use Terms | Privacy Source.