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Publication : Simple and large-scale chromosomal engineering of mouse zygotes via in vitro and in vivo electroporation.

First Author  Iwata S Year  2019
Journal  Sci Rep Volume  9
Issue  1 Pages  14713
PubMed ID  31604975 Mgi Jnum  J:284649
Mgi Id  MGI:6389444 Doi  10.1038/s41598-019-50900-y
Citation  Iwata S, et al. (2019) Simple and large-scale chromosomal engineering of mouse zygotes via in vitro and in vivo electroporation. Sci Rep 9(1):14713
abstractText  The clustered regularly interspaced short palindromic repeats (CRISPR)/Cas9 system has facilitated dramatic progress in the field of genome engineering. Whilst microinjection of the Cas9 protein and a single guide RNA (sgRNA) into mouse zygotes is a widespread method for producing genetically engineered mice, in vitro and in vivo electroporation (which are much more convenient strategies) have recently been developed. However, it remains unknown whether these electroporation methods are able to manipulate genomes at the chromosome level. In the present study, we used these techniques to introduce chromosomal inversions of several megabases (Mb) in length in mouse zygotes. Using in vitro electroporation, we successfully introduced a 7.67 Mb inversion, which is longer than any previously reported inversion produced using microinjection-based methods. Additionally, using in vivo electroporation, we also introduced a long chromosomal inversion by targeting an allele in F1 hybrid mice. To our knowledge, the present study is the first report of target-specific chromosomal inversions in mammalian zygotes using electroporation.
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