Frequent loss of heterozygosity in CRISPR-Cas9-edited early human embryos.
Proc Natl Acad Sci U S A
; 118(22)2021 06 01.
Article
em En
| MEDLINE
| ID: mdl-34050011
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 â¼16% 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.
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Texto completo:
1
Base de dados:
MEDLINE
Assunto principal:
Blastocisto
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Perda de Heterozigosidade
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Fator 3 de Transcrição de Octâmero
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Sistemas CRISPR-Cas
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Células-Tronco Embrionárias Humanas
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Edição de Genes
Idioma:
En
Ano de publicação:
2021
Tipo de documento:
Article
País de afiliação:
Reino Unido