Whole chromosome loss and genomic instability in mouse embryos after CRISPR-Cas9 genome editing

• Published in: Nature communications Vol. 12; no. 1; p. 5855
• Main Authors: Papathanasiou, Stamatis, Markoulaki, Styliani, Blaine, Logan J., Leibowitz, Mitchell L., Zhang, Cheng-Zhong, Jaenisch, Rudolf, Pellman, David
• Format: Journal Article
• Language: English
• Published: London Nature Publishing Group UK 06-10-2021; Nature Publishing Group; Nature Portfolio
• Subjects: 14/63; 45/23; 631/208/211; 631/208/4041/3196; 631/80/641/2002; 64/60; Animals; Chromosome Segregation; Chromosome Structures; Chromosomes; Complications; CRISPR; CRISPR-Cas Systems; Editing; Embryo, Mammalian; Embryogenesis; Embryonic Development - genetics; Embryonic growth stage; Embryos; Gene Editing - methods; Gene sequencing; Genome editing; Genomes; Genomic Instability; Humanities and Social Sciences; Karyotype; Karyotypes; Mice; Micronuclei; Mitosis; multidisciplinary; Science; Science (multidisciplinary); Side effects; Whole Genome Sequencing
• ISSN: 2041-1723; 2041-1723
• DOI: 10.1038/s41467-021-26097-y

Karyotype alterations have emerged as on-target complications from CRISPR-Cas9 genome editing. However, the events that lead to these karyotypic changes in embryos after Cas9-treatment remain unknown. Here, using imaging and single-cell genome sequencing of 8-cell stage embryos, we track both spontaneous and Cas9-induced karyotype aberrations through the first three divisions of embryonic development. We observe the generation of abnormal structures of the nucleus that arise as a consequence of errors in mitosis, including micronuclei and chromosome bridges, and determine their contribution to common karyotype aberrations including whole chromosome loss that has been recently reported after editing in embryos. Together, these data demonstrate that Cas9-mediated germline genome editing can lead to unwanted on-target side effects, including major chromosome structural alterations that can be propagated over several divisions of embryonic development.
A possible undesired outcome of CRISPR-Cas9 germline editing is unwanted karyotype alterations. Here the authors track aberrations through three divisions of embryonic development following Cas9 editing.