Abstract
Genome editing has potential for the targeted correction of germline mutations. Here we describe the correction of the heterozygous MYBPC3 mutation in human preimplantation embryos with precise CRISPR-Cas9-based targeting accuracy and high homology-directed repair efficiency by activating an endogenous, germline-specific DNA repair response. Induced double-strand breaks (DSBs) at the mutant paternal allele were predominantly repaired using the homologous wild-type maternal gene instead of a synthetic DNA template. By modulating the cell cycle stage at which the DSB was induced, we were able to avoid mosaicism in cleaving embryos and achieve a high yield of homozygous embryos carrying the wild-type MYBPC3 gene without evidence of off-target mutations. The efficiency, accuracy and safety of the approach presented suggest that it has potential to be used for the correction of heritable mutations in human embryos by complementing preimplantation genetic diagnosis. However, much remains to be considered before clinical applications, including the reproducibility of the technique with other heterozygous mutations.
| Original language | English |
|---|---|
| Pages (from-to) | 413-419 |
| Number of pages | 7 |
| Journal | Nature |
| Volume | 548 |
| Issue number | 7668 |
| DOIs | |
| State | Published - 24 Aug 2017 |
Bibliographical note
Funding Information:Acknowledgements We acknowledge the OHSU Institutional Review Board (IRB), Innovative Research Advisory Panel (IRAP), Scientific Review Committee (SRC) and Data Safety Monitoring Committee (DSMC) for oversight and guidance on this study. We thank all study participants for tissue donations; the Women’s Health Research Unit staff, IVF laboratory staff, University Fertility Consultants and the Reproductive Endocrinology and Infertility Division in the Department of Obstetrics and Gynecology, OHSU for support and procurement of human gametes; S. Olson and Research Cytogenetics Laboratory at OHSU for cytogenetic analysis of ES cells; S. Cooper from the Wallace Division of Smiths Medical for donating ICSI and gamete manipulation micropipettes; Y. Wang, T. Wu and Y. Shen from BGI-Shenzhen for help with sample preparation, sequencing and data analyses; M. Ku from the H. A. and Mary K. Chapman Charitable Foundations Genomic Sequencing Core of the Salk Institute for next generation sequencing; and E. Aizawa and R. Hernandez-Benitez from the laboratory of JCIB for assistance. Studies conducted at OHSU were supported by OHSU institutional funds. Work in the laboratory of J.-S.K. was supported by the Institute for Basic Science (IBS-R021-D1). Work in the laboratory of J.C.I.B. was supported by the G. Harold and Leila Y. Mathers Charitable Foundation, the Moxie Foundation and the Leona M. and Harry B. Helmsley Charitable Trust. Work at BGI was supported by the Shenzhen Municipal Government of China (DRC-SZ [2016] 884).
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