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

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 similar to 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. not only an indispensable molecular biology technique (1) but also has enormous therapeutic potential as a tool to correct disease-causing mutations (2). Genome editing of human embryos or germ cells to produce heritable changes has the potential to reduce the burden of genetic disease, and its use in this context is currently a topic of international discussions centered around ethics, safety, and efficiency (3, 4).

Automated summary

CRISPR-Cas9 genome editing shows promise for correcting disease-associated alleles in somatic cells, but studies in human embryos reveal challenges such as low mutation repair efficiency, high mosaicism, and unintended editing outcomes. Further research is crucial to evaluate the safety of genome editing techniques in human embryos.