説明
<jats:title>Abstract</jats:title><jats:p>The use of CRISPR-Cas9 as a therapeutic reagent is hampered by its off-target effects. Although rationally designed <jats:italic>S. pyogenes</jats:italic> 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 develop a directed evolution approach in <jats:italic>E. coli</jats:italic> to obtain Sniper-Cas9, which shows high specificities without killing on-target activities in human cells. Unlike other engineered Cas9 variants, Sniper-Cas9 shows WT-level on-target activities with extended or truncated sgRNAs with further reduced off-target activities and works well in a preassembled ribonucleoprotein (RNP) format to allow DNA-free genome editing.</jats:p>
収録刊行物
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- Nature Communications
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Nature Communications 9 (1), 3048-, 2018-08-06
Springer Science and Business Media LLC