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Generation of CRISPR/Cas9-mediated bicistronic knock-in ins1-cre driver mice

In the present study, we generated novel cre driver mice for gene manipulation in pancreatic β cells. Using the CRISPR/Cas9 system, stop codon sequences of Ins1 were targeted for insertion of cre, including 2A sequences. A founder of C57BL/6J-Ins1(em1 (cre) Utr) strain was produced from an oocyte in...

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Detalles Bibliográficos
Autores principales: Hasegawa, Yoshikazu, Hoshino, Yoshikazu, Ibrahim, Abdelaziz E., Kato, Kanako, Daitoku, Yoko, Tanimoto, Yoko, Ikeda, Yoshihisa, Oishi, Hisashi, Takahashi, Satoru, Yoshiki, Atsushi, Yagami, Ken-ichi, Iseki, Hiroyoshi, Mizuno, Seiya, Sugiyama, Fumihiro
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Japanese Association for Laboratory Animal Science 2016
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4976246/
https://www.ncbi.nlm.nih.gov/pubmed/27053096
http://dx.doi.org/10.1538/expanim.16-0016
Descripción
Sumario:In the present study, we generated novel cre driver mice for gene manipulation in pancreatic β cells. Using the CRISPR/Cas9 system, stop codon sequences of Ins1 were targeted for insertion of cre, including 2A sequences. A founder of C57BL/6J-Ins1(em1 (cre) Utr) strain was produced from an oocyte injected with pX330 containing the sequences encoding gRNA and Cas9 and a DNA donor plasmid carrying 2A-cre. (R26GRR x C57BL/6J-Ins1(em1 (cre) Utr)) F(1) mice were histologically characterized for cre-loxP recombination in the embryonic and adult stages; cre-loxP recombination was observed in all pancreatic islets examined in which almost all insulin-positive cells showed tdsRed fluorescence, suggesting β cell-specific recombination. Furthermore, there were no significant differences in results of glucose tolerance test among genotypes (homo/hetero/wild). Taken together, these observations indicated that C57BL/6J-Ins1(em1 (cre) Utr) is useful for studies of glucose metabolism and the strategy of bicistronic cre knock-in using the CRISPR/Cas9 system could be useful for production of cre driver mice.