March 28th, 2013 | Category: Papers, Protein design, Synthetic Biology | Leave a comment

Cas9 – a new genome editing tool

Cas9 – could it change Biotech forever? Well, you’ll have to follow that link to find out. In the meantime, here’s a very brief introduction.

CRISPR – clustered, regularly interspersed, short palindromic repeats – are used by bacteria and archaea to provide protection against foreign nucleic acid sequences. CRISPR-mediated immune systems use CRISPR-associated (Cas) endonucleases that are directed to the target DNA by two small RNA sequences. This provides  sequence-specific detection and silencing of foreign nucleic acids (Wiedenheft et al. 2012). Last year, Jinek et al. (2012) demonstrated that Cas9 from Streptococcus pyogenes could be programmed to target a desired DNA sequence in vitro. Now, within nine months, six further papers have exploited this system to edit precisely the genomes of a range of different species in vivo. These include human cell lines, zebrafish embryos, Streptococcus pyogenes, S. pneumoniae, Saccharomyces cerevisiae and E. coli. More detailed explanations of how the system works can be found here (Nature Methods) and of course in the papers themselves (which are multiplying as I type). Given the speed of uptake of new technology in Biotech. and synthetic biology circles, I can’t help but think this will be appearing in one or two iGEM projects this year.

Here are the papers, in no particular order:

Cong et al. (2013) Multiplex Genome Engineering Using CRISPR/Cas Systems Science;

Cho et al. (2013) Targeted genome engineering in human cells with the Cas9 RNA-guided endonuclease Nature Biotechnology.;

Mali et al. (2013) RNA-Guided Human Genome Engineering via Cas9 Science;

Jiang et al. (2013) RNA-guided editing of bacterial genomes using CRISPR-Cas systems Nature Biotechnology;

Hwang et al. (2013) Efficient genome editing in zebrafish using a CRISPR-Cas system Nature Biotechnology;

DiCarlo et al. (2013) Genome engineering in Saccharomyces cerevisiae using CRISPR-Cas systems Nucleic Acids Research.

Update:

Shen et al. (2013) Generation of gene-modified mice via Cas9/RNA-mediated gene targeting Cell Research

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