bioRxivBiochemistry
Directed evolution of CRISPR-Cas9 to increase its specificity
J. K. Lee, E. Jeong, J. Lee, M. Jung, E. Shin, Y.-h. Kim, K. Lee, D. Kim, J.-S. Kim, S. Kim
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.
- organism: E. coli
- organism: human
- assay: CRISPR screen
- research focus: protein design
- approach: experimental assay
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