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xCas9: Engineering a CRISPR Variant with PAM Flexibility

Posted by Mary Gearing on Mar 28, 2018 2:52:13 PM

In order to bind DNA, Cas9 and other CRISPR enzymes require a short PAM sequence adjacent to the targeted sequence at the locus of interest. SpCas9’s 3’ NGG PAM occurs frequently in GC-rich genomes, but a PAM is not always available near the locus you’d like to modify. To tackle the PAM problem, researchers have engineered alternative Cas9s binding distinct PAM sequences. Now, Hu et al., working in David Liu’s lab, have gone one step further, using directed evolution to create xCas9, an enzyme recognizing a broad range of PAMs like NG, GAA, and GAT, but also displaying increased editing specificity. We’re excited to learn more about xCas9 - here’s what we know so far!

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Topics: CRISPR

CRISPR 101: Cas9 Nickase Design and Homology Directed Repair

Posted by Mary Gearing on Mar 15, 2018 8:59:40 AM

By mutating one of two Cas9 nuclease domains, researchers created the CRISPR nickase. Nickases create a single-strand rather than a double-strand break, and when used with two adjacent gRNAs, can lower the probability of off-target editing. But that’s not all! New research from IDT (Integrated DNA Technologies) has shown that a nickase approach can improve homology directed repair (HDR) rates, provided you follow some simple design rules described below.

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Topics: CRISPR, CRISPR 101

CRISPR Challenges: Standardization and Homology Directed Repair

Posted by Mary Gearing on Feb 14, 2018 9:36:24 AM

Blugene and I represented Addgene at the recent Keystone meeting on Precision Genome Editing with Programmable Nucleases. Check out #KSgenome on Twitter if you missed our live updates!

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Topics: CRISPR

Genome Engineering, CRISPR, and Research in Singapore: Interview with Wei Leong Chew

Posted by Tyler Ford on Feb 6, 2018 9:00:56 AM

In today’s podcast, we sit down with Wei Leong Chew, a researcher at the Genome Institute of Singapore who recently started his own lab. We discuss some of the joys and difficulties of getting a lab up and running, and learn a little bit about what it was like for Wei Leong to work in George Church’s lab as a graduate student.

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Topics: Genome Engineering, CRISPR, Podcast

New CRISPR Web Resources and #12DaysofCRISPR Recap

Posted by Mary Gearing on Dec 12, 2017 10:13:34 AM

Since the start of the CRISPR revolution, Addgene has distributed over 100,000 CRISPR plasmids. But that’s not our only job - we strive to also give you high-quality educational resources to help you do better research. CRISPR is an incredibly fast-moving field, and we want to make it easy for you to keep up with new developments (and, of course, find plasmids that will be useful to you.)

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Topics: CRISPR, CRISPR 101, Using Addgene's Website

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