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Animal Bioresource in Japan

.: Home > Animal Bioresource in Japan > 2014 > Volume 63 Number 2 > Yoshikazu Hasegawa, Yoko Daitoku, Seiya Mizuno, Yoko Tanimoto, Saori Mizuno-Iijima, Miki Matsuo, Noriko Kajiwara, Masatsugu Ema, Hisashi Oishi, Yoshihiro Miwa, Kazuyuki Mekada, Atsushi Yoshiki, Satoru Takahashi, Fumihiro Sugiyama, Ken-ichi Yagami

Generation and Characterization of Ins1-cre-driver C57BL/6N for Exclusive Pancreatic Beta Cell-specific Cre-loxP Recombination

Yoshikazu Hasegawa, Yoko Daitoku, Seiya Mizuno, Yoko Tanimoto, Saori Mizuno-Iijima, Miki Matsuo, Noriko Kajiwara, Masatsugu Ema, Hisashi Oishi, Yoshihiro Miwa, Kazuyuki Mekada, Atsushi Yoshiki, Satoru Takahashi, Fumihiro Sugiyama, Ken-ichi Yagami
Laborarory Animal Resource Center, University of Tsukuba, 1-1-1 Tennodai, Tsukuba 305-8575, Japan
Abstract :

Cre/loxP system-mediated site-specific recombination is utilized to study gene function in vivo. Successful conditional knockout of genes of interest is dependent on the availability of Cre-driver mice. We produced and characterized pancreatic β cell-specific Cre-driver mice for use in diabetes mellitus research. The gene encoding Cre was inserted into the second exon of mouse Ins1 in a bacterial artificial chromosome (BAC). Five founder mice were produced by microinjection of linearized BAC Ins1-cre. The transgene was integrated between Mafa and the telomere on chromosome 15 in one of the founders, BAC Ins1-cre25. To investigate Cre-loxP recombination, BAC Ins1-cre25 males were crossed with two different Cre-reporters, R26R and R26GRR females. On gross observation, reporter signal after Cre-loxP recombination was detected exclusively in the adult pancreatic islets in both F1 mice. Immunohistological analysis indicated that Cre-loxP recombination-mediated reporter signal was colocalized with insulin in pancreatic islet cells of both F1 mice, but not with glucagon. Moreover, Cre-loxP recombination signal was already observed in the pancreatic islets at E13.5 in both F1 fetuses. Finally, we investigated ectopic Cre-loxP recombination for Ins1, because the ortholog Ins2 is also expressed in the brain, in addition to the pancreas. However, there was no Cre-loxP recombination-mediated reporter signal in the brain of both F1 mice. Our data suggest that BAC Ins1-cre25 mice are a useful Cre-driver C57BL/6N for pancreatic β cell-specific Cre-loxP recombination, except for crossing with knock-in mice carrying floxed gene on chromosome 15.

Keywords :
cre-driver mice, cre-loxP recombination, diabetes, insulin1, pancreatic β cells

Date Deposited : 07 Apr 2015 10:05

Last Modified : 07 Apr 2015 10:05

Official URL: http://https://www.jstage.jst.go.jp/browse/expanim/62/1/_contents

Volume 63, Number 2, - 2014

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