By Mary Gearing
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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 ...
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. In this post, we’ll summarize how IDT ...
This post was updated on Jul 27, 2020. CRISPR, and specifically Cas9 from S. pyogenes (SpCas9), is truly an exceptional genome engineering tool. It is easy to use, functional in most species, and has many applications. That said, SpCas9 is not the only game in town, and other ...
Cas9 is the genome editing tool of choice for a number of model organisms: mammalian cells, yeast, drosophila, plants, worms, zebrafish, frogs, some bacteria; but not thermophilic (high heat loving) bacteria. Until recently the only available Cas9 proteins were isolated from ...
Note: Cpf1 is also called Cas12a. There’s a new development for CRISPR-Cpf1 genome editing! A recent paper from Feng Zhang's lab describes how to use Cpf1 for multiplex genome editing. For a few reasons, Cpf1 is a simplified system for editing multiple targets compared to ...
Last updated Oct 7, 2020 by Gabrielle Clouse. This post was contributed by guest bloggers Marcelle Tuttle and Alex Chavez, researchers at the Wyss Institute for Biologically Inspired Engineering. CRISPR/Cas9 is an enormously plastic tool and has taken the scientific world by ...
Update (November 18, 2016): Researchers from a variety of institutions recently reported their inability to recapitulate the results of Gao et al 2016 in a letter to Protein & Cell. Update (August 3rd, 2017) THE ORIGINAL NgAgo ARTICLE DISCUSSED IN THIS POST HAS BEEN RETRACTED ...
