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Search: WFRF:(Chan BKC)

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  • Thean, DGL, et al. (author)
  • Machine learning-coupled combinatorial mutagenesis enables resource-efficient engineering of CRISPR-Cas9 genome editor activities
  • 2022
  • In: Nature communications. - : Springer Science and Business Media LLC. - 2041-1723. ; 13:1, s. 2219-
  • Journal article (peer-reviewed)abstract
    • The genome-editing Cas9 protein uses multiple amino-acid residues to bind the target DNA. Considering only the residues in proximity to the target DNA as potential sites to optimise Cas9’s activity, the number of combinatorial variants to screen through is too massive for a wet-lab experiment. Here we generate and cross-validate ten in silico and experimental datasets of multi-domain combinatorial mutagenesis libraries for Cas9 engineering, and demonstrate that a machine learning-coupled engineering approach reduces the experimental screening burden by as high as 95% while enriching top-performing variants by ∼7.5-fold in comparison to the null model. Using this approach and followed by structure-guided engineering, we identify the N888R/A889Q variant conferring increased editing activity on the protospacer adjacent motif-relaxed KKH variant of Cas9 nuclease from Staphylococcus aureus (KKH-SaCas9) and its derived base editor in human cells. Our work validates a readily applicable workflow to enable resource-efficient high-throughput engineering of genome editor’s activity.
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  • Result 1-3 of 3
Type of publication
journal article (3)
Type of content
peer-reviewed (3)
Author/Editor
Zheng, ZL (3)
Choi, GCG (3)
Zhou, P (3)
Chan, BKC (3)
Wong, ASL (3)
Tan, K. (2)
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Yuen, CTL (2)
Chu, HY (2)
Wong, KH (2)
Mak, SYL (2)
Li, XR (1)
Xu, F (1)
Bao, SY (1)
Thean, D (1)
Chan, HYE (1)
Chow, MZY (1)
Thean, DGL (1)
Fong, JHC (1)
Kwok, CCS (1)
Chan, YM (1)
Ho, JWK (1)
Wan, YK (1)
Tong, CSW (1)
Zhong, SSW (1)
Sun, JR (1)
Bao, YF (1)
Khaw, JV (1)
Leung, SY (1)
Cheung, LWT (1)
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University
Karolinska Institutet (3)
Language
English (3)

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