Repository logo
 

Frequent loss of heterozygosity in CRISPR-Cas9-edited early human embryos.

Accepted version
Peer-reviewed

No Thumbnail Available

Type

Article

Change log

Authors

Alanis-Lobato, Gregorio 
McCarthy, Afshan 
Kubikova, Nada 

Abstract

CRISPR-Cas9 genome editing is a promising technique for clinical applications, such as the correction of disease-associated alleles in somatic cells. The use of this approach has also been discussed in the context of heritable editing of the human germ line. However, studies assessing gene correction in early human embryos report low efficiency of mutation repair, high rates of mosaicism, and the possibility of unintended editing outcomes that may have pathologic consequences. We developed computational pipelines to assess single-cell genomics and transcriptomics datasets from OCT4 (POU5F1) CRISPR-Cas9-targeted and control human preimplantation embryos. This allowed us to evaluate on-target mutations that would be missed by more conventional genotyping techniques. We observed loss of heterozygosity in edited cells that spanned regions beyond the POU5F1 on-target locus, as well as segmental loss and gain of chromosome 6, on which the POU5F1 gene is located. Unintended genome editing outcomes were present in ∼16% of the human embryo cells analyzed and spanned 4-20 kb. Our observations are consistent with recent findings indicating complexity at on-target sites following CRISPR-Cas9 genome editing. Our work underscores the importance of further basic research to assess the safety of genome editing techniques in human embryos, which will inform debates about the potential clinical use of this technology.

Description

Keywords

CRISPR-Cas9, genome editing, human embryo, loss of heterozygosity, segmental aneuploidy, Blastocyst, CRISPR-Cas Systems, Cell Line, Chromosomes, Human, Pair 6, Gene Editing, Human Embryonic Stem Cells, Humans, Loss of Heterozygosity, Octamer Transcription Factor-3

Journal Title

Proc Natl Acad Sci U S A

Conference Name

Journal ISSN

0027-8424
1091-6490

Volume Title

118

Publisher

Proceedings of the National Academy of Sciences

Rights

All rights reserved
Sponsorship
Wellcome Trust (221856/Z/20/Z)