Abstract:
:Although genetically modified mice can be generated with high efficiency by using CRISPR/Cas9-mediated genome editing in mouse zygotes, only the loci with a protospacer-adjacent motif (PAM) sequence are targetable. The present study investigated the usability of engineered Streptococcus pyogenes Cas9 (SpCas9-NG) in mouse zygotes. In addition to the 5'-NGG sequence, SpCas9-NG recognized the 5'-NGA, 5'-NGC and 5'-NGT sequences in mouse zygotes as PAMs that were appropriate for the generation of knockout mice. Moreover, SpCas9-NG-mediated genome editing enabled the generation of knock-in mice untargetable by the conventional SpCas9 in mouse zygotes. These results suggest that SpCas9-NG-mediated genome editing in zygotes is available for the generation of knockout and knock-in mice at the locus corresponding to NGN-PAM.
journal_name
Sci Repjournal_title
Scientific reportsauthors
Fujii W,Ito H,Kanke T,Ikeda A,Sugiura K,Naito Kdoi
10.1038/s41598-019-49394-5subject
Has Abstractpub_date
2019-09-09 00:00:00pages
12878issue
1issn
2045-2322pii
10.1038/s41598-019-49394-5journal_volume
9pub_type
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