RT Journal Article
SR Electronic
T1 FREQUENT GENE CONVERSION IN HUMAN EMBRYOS INDUCED BY DOUBLE STRAND BREAKS
JF bioRxiv
FD Cold Spring Harbor Laboratory
SP 2020.06.19.162214
DO 10.1101/2020.06.19.162214
A1 Dan Liang
A1 Nuria Marti Gutierrez
A1 Tailai Chen
A1 Yeonmi Lee
A1 Sang-Wook Park
A1 Hong Ma
A1 Amy Koski
A1 Riffat Ahmed
A1 Hayley Darby
A1 Ying Li
A1 Crystal Van Dyken
A1 Aleksei Mikhalchenko
A1 Thanasup Gonmanee
A1 Tomonari Hayama
A1 Han Zhao
A1 Keliang Wu
A1 Jingye Zhang
A1 Zhenzhen Hou
A1 Jumi Park
A1 Chong-Jai Kim
A1 Jianhui Gong
A1 Yilin Yuan
A1 Ying Gu
A1 Yue Shen
A1 Susan B. Olson
A1 Hui Yang
A1 David Battaglia
A1 Thomas O’Leary
A1 Sacha A. Krieg
A1 David M. Lee
A1 Diana H. Wu
A1 P. Barton Duell
A1 Sanjiv Kaul
A1 Jin-Soo Kim
A1 Stephen B. Heitner
A1 Eunju Kang
A1 Zi-Jiang Chen
A1 Paula Amato
A1 Shoukhrat Mitalipov
YR 2020
UL http://biorxiv.org/content/early/2020/06/20/2020.06.19.162214.abstract
AB Applications of genome editing ultimately depend on DNA repair triggered by targeted double-strand breaks (DSBs). However, repair mechanisms in human cells remain poorly understood and vary across different cell types. Here we report that DSBs selectively induced on a mutant allele in heterozygous human embryos are repaired by gene conversion using an intact wildtype homolog as a template in up to 40% of targeted embryos. We also show that targeting of homozygous loci facilitates an interplay of non-homologous end joining (NHEJ) and gene conversion and results in embryos which carry identical indel mutations on both loci. Additionally, conversion tracks may expand bidirectionally well beyond the target region leading to an extensive loss of heterozygosity (LOH). Our study demonstrates that gene conversion and NHEJ are two major DNA DSB repair mechanisms in preimplantation human embryos. While gene conversion could be applicable for gene correction, extensive LOH presents a serious safety concern.Competing Interest StatementThe authors have declared no competing interest.