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Sep 12, 2013
 |  BC Innovations  |  Tools & Techniques

Crisper results for CRISPR

Since its introduction earlier this year, proponents of CRISPR-based genome editing have touted its simplicity and efficiency over other genome editing platforms, but they have more recently also offered a word of caution on its specificity.1-3 Research from the Broad Institute of MIT and Harvard may now allay these concerns with an updated version of the technique that decreases editing activity at predicted off-target sites by 50- to 1,000-fold compared with earlier implementations.4

The group now needs to assess the approach's off-target editing activity on the whole-genome level.

CRISPR (clustered, regularly interspaced short palindromic repeats)-based genome editing, and more specifically CRISPR-Cas9 (CRISPR-associated protein 9)-based genome editing, came into the limelight this January after multiple research groups showed proof of concept for engineering site-specific mutations in the genomes of key model organisms including bacterial and mammalian cells, zebrafish and rodents.5-11

Less than four months after this initial flurry of results, one of the groups published the use of the CRISPR-Cas9 system to simultaneously engineer point mutations into multiple target genes in a mouse,12,13 which typically is a time- and labor-intensive process that usually would require several rounds of interbreeding.

The platform is derived from an acquired immunity-like system in bacteria, whereby short CRISPR-encoded RNAs guide CRISPR-associated proteins such as the Cas9 endonuclease to cleave homologous foreign DNA contained within plasmids or bacteriophages.

In the CRISPR-Cas9 platform, the specificity of the Cas9 endonuclease is dictated by base pairing between a custom-designed single guide RNA (sgRNA) and the target DNA sequence on the...

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