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bioRxiv · 10.1101/237040

Directed evolution of CRISPR-Cas9 to increase its specificity

Lee, J. K.·Jeong, E.·Lee, J.·Jung, M.·Shin, E.·Kim, Y.-h.·Lee, K.·Kim, D.·Kim, J.-S.·Kim, S.
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Abstract

The use of CRISPR-Cas9 as a therapeutic reagent is hampered by its off-target effects. Although rationally designed S. pyogenes Cas9 (SpCas9) variants that display higher specificities than the wild-type SpCas9 protein are available, these attenuated Cas9 variants are often poorly efficient in human cells. Here, we have used a directed evolution approach in E. coli to obtain Sniper-Cas9, which shows high specificities without sacrificing on-target activities in human cells.

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Lee, J. K., Jeong, E., Lee, J., Jung, M., Shin, E., Kim, Y.-h., Lee, K., Kim, D., Kim, J.-S., Kim, S.. 2017-12-19. Directed evolution of CRISPR-Cas9 to increase its specificity. https://doi.org/10.1101/237040

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