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유전체교정 연구단
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Directed evolution of CRISPR-Cas9 to increase its specificity

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Title
Directed evolution of CRISPR-Cas9 to increase its specificity
Author(s)
Jungjoon K. Lee; Euihwan Jeong; Joonsun Lee; Minhee Jung; Eunji Shin; Young-hoon Kim; Kangin Lee; Inyoung Jung; Daesik Kim; Seokjoong Kim; Jin-Soo Kim
Publication Date
2018-08
Journal
NATURE COMMUNICATIONS, v.9, no.1, pp.3048 -
Publisher
NATURE PUBLISHING GROUP
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 develop a directed evolution approach in E. coli 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. © 2018, The Author(s)
URI
https://pr.ibs.re.kr/handle/8788114/5533
ISSN
2041-1723
Appears in Collections:
Center for Genome Engineering(유전체 교정 연구단) > Journal Papers (저널논문)
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