Abstract | ||
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Following Cas9 cleavage, DNA repair without a donor template is generally considered stochastic, heterogeneous and impractical beyond gene disruption. Here, we show that template-free Cas9 editing is predictable and capable of precise repair to a predicted genotype, enabling correction of disease-associated mutations in humans. We constructed a library of 2,000 Cas9 guide RNAs paired with DNA target sites and trained inDelphi, a machine learning model that predicts genotypes and frequencies of 1-to 60-base-pair deletions and 1-base-pair insertions with high accuracy (r = 0.87) in five human and mouse cell lines. inDelphi predicts that 5-11% of Cas9 guide RNAs targeting the human genome are 'precise-50', yielding a single genotype comprising greater than or equal to 50% of all major editing products. We experimentally confirmed precise-50 insertions and deletions in 195 human disease-relevant alleles, including correction in primary patient-derived fibroblasts of pathogenic alleles to wild-type genotype for Hermansky-Pudlak syndrome and Menkes disease. This study establishes an approach for precise, template-free genome editing. |
Year | DOI | Venue |
---|---|---|
2018 | 10.1038/s41586-018-0686-x | NATURE |
DocType | Volume | Issue |
Journal | 563 | 7733 |
ISSN | Citations | PageRank |
0028-0836 | 0 | 0.34 |
References | Authors | |
1 | 10 |
Name | Order | Citations | PageRank |
---|---|---|---|
Max Shen | 1 | 11 | 2.35 |
Mandana Arbab | 2 | 0 | 0.34 |
Jonathan Y Hsu | 3 | 0 | 0.34 |
Daniel Worstell | 4 | 0 | 0.68 |
Sannie J Culbertson | 5 | 0 | 0.34 |
Olga Krabbe | 6 | 0 | 0.34 |
Christopher A. Cassa | 7 | 7 | 3.32 |
David R Liu | 8 | 0 | 0.34 |
David K. Gifford | 9 | 2956 | 946.76 |
Richard I Sherwood | 10 | 0 | 1.01 |