Revealing hidden complexities of genomic rearrangements generated with Cas9.
Publication Date
2017-10-09Journal Title
Sci Rep
ISSN
2045-2322
Publisher
Springer Science and Business Media LLC
Volume
7
Issue
1
Pages
12867
Language
eng
Type
Article
This Version
VoR
Physical Medium
Electronic
Metadata
Show full item recordCitation
Boroviak, K., Fu, B., Yang, F., Doe, B., & Bradley, A. (2017). Revealing hidden complexities of genomic rearrangements generated with Cas9.. Sci Rep, 7 (1), 12867. https://doi.org/10.1038/s41598-017-12740-6
Abstract
Modelling human diseases caused by large genomic rearrangements has become more accessible since the utilization of CRISPR/Cas9 in mammalian systems. In a previous study, we showed that genomic rearrangements of up to one million base pairs can be generated by direct injection of CRISPR/Cas9 reagents into mouse zygotes. Although these rearrangements are ascertained by junction PCR, we describe here a variety of anticipated structural changes often involving reintegration of the region demarcated by the gRNAs in the vicinity of the edited locus. We illustrate here some of this diversity detected by high-resolution fibre-FISH and conclude that extensive molecular analysis is required to fully understand the structure of engineered chromosomes generated by Cas9.
Keywords
Animals, Mice, Inbred C57BL, In Situ Hybridization, Fluorescence, Gene Rearrangement, Gene Duplication, Sequence Deletion, Genome, Female, Sequence Inversion, NADPH Oxidase 4, CRISPR-Associated Protein 9
Identifiers
External DOI: https://doi.org/10.1038/s41598-017-12740-6
This record's URL: https://www.repository.cam.ac.uk/handle/1810/311501
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