RRC ID 54139
Author Hasegawa Y, Hoshino Y, Ibrahim AE, Kato K, Daitoku Y, Tanimoto Y, Ikeda Y, Oishi H, Takahashi S, Yoshiki A, Yagami K, Iseki H, Mizuno S, Sugiyama F.
Title Generation of CRISPR/Cas9-mediated bicistronic knock-in ins1-cre driver mice.
Journal Exp Anim
Abstract 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)) F1 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.
Volume 65(3)
Pages 319-27
Published 2016-7-29
DOI 10.1538/expanim.16-0016
PMID 27053096
PMC PMC4976246
MeSH Animals CRISPR-Cas Systems* Codon, Terminator / genetics Crk-Associated Substrate Protein / administration & dosage Gene Editing / methods* Glucose / metabolism Injections Insulin / genetics* Insulin-Secreting Cells* Integrases / administration & dosage Integrases / genetics* Mice, Inbred C57BL Mice, Mutant Strains* / genetics Mutagenesis, Insertional Oocytes RNA / administration & dosage Recombination, Genetic
IF 1.574
Times Cited 7
DNA material px330-Ins1 (RDB13945) pIns1/2A-cre (RDB13946) p2color-Ins1 (RDB13947) p2color (RDB13948)