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Characterizing CRISPR off-target effects

Nature Reviews Genetics volume 15, page 5 (2014)Cite this article

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Off-target mutagenesis is an emerging problem when inducing site-specific DNA breaks for genome editing. Cho et al. used high-throughput DNA sequencing to characterize off-target mutagenesis in human cells during editing by the RNA-guided CRISPR–Cas system. They found that off-target mutagenesis occurs frequently at sites that differ by one nucleotide from the intended target site but substantially less frequently at sites with more than one mismatch. Off-target mutagenesis could be reduced through careful design of the guide RNA sequence and by using alternative nucleases to induce paired single-strand rather than double-strand DNA breaks.

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References

  1. Cho, S. W. et al. Analysis of off-target effects of CRISPR/Cas-derived RNA-guided endonucleases and nickases. Genome Res. http://www.dx.doi.org/10.1101/gr.162339.113 (2013)

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  1. Darren J. Burgess
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Burgess, D. Characterizing CRISPR off-target effects. Nat Rev Genet 15, 5 (2014). https://doi.org/10.1038/nrg3651

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