RRC ID 73461
Author Hasegawa Y, Daitoku Y, Sekiguchi K, Tanimoto Y, Mizuno-Iijima S, Mizuno S, Kajiwara N, Ema M, Miwa Y, Mekada K, Yoshiki A, Takahashi S, Sugiyama F, Yagami K.
Title Novel ROSA26 Cre-reporter knock-in C57BL/6N mice exhibiting green emission before and red emission after Cre-mediated recombination.
Journal Exp Anim
Abstract The Cre/loxP system is a strategy for controlling temporal and/or spatial gene expression through genome alteration in mice. As successful Cre/loxP genome alteration depends on Cre-driver mice, Cre-reporter mice are essential for validation of Cre gene expression in vivo. In most Cre-reporter mouse strains, although the presence of reporter product indicates the expression of Cre recombinase, it has remained unclear whether a lack of reporter signal indicates either no Cre recombinase expression or insufficient reporter gene promoter activity. We produced a novel ROSA26 knock-in Cre-reporter C57BL/6N strain exhibiting green emission before and red after Cre-mediated recombination, designated as strain R26GRR. Ubiquitous green fluorescence and no red fluorescence were observed in R26GRR mice. To investigate the activation of tdsRed, EGFP-excised R26GRR, R26RR, mice were produced through the crossing of C57BL/6N mice with R26GRR/Ayu1-Cre F1 mice. R26RR mice showed extraordinarily strong red fluorescence in almost all tissues examined, suggesting ubiquitous activation of the second reporter in all tissues after Cre/loxP recombination. Moreover, endothelial cell lineage and pancreatic islet-specific expression of red fluorescence were detected in R26GRR/Tie2-Cre F1 mice and R26GRR /Ins1-Cre F1 mice, respectively. These results indicated that R26GRR mice are a useful novel Cre-reporter mouse strain. In addition, R26GRR mice with a pure C57BL/6N background represent a valuable source of green-to-red photoconvertible cells following Cre/loxP recombination for application in transplantation studies. The R26GRR mouse strain will be available from RIKEN BioResource Center (http://www.brc.riken.jp/lab/animal/en/).
Volume 62(4)
Pages 295-304
Published 2013-1-1
DOI 10.1538/expanim.62.295
PII DN/JST.JSTAGE/expanim/62.295
PMID 24172193
PMC PMC4160954
MeSH Animals Cells, Cultured Embryonic Stem Cells Endothelial Cells / metabolism Female Gene Expression* Gene Knock-In Techniques Genes, Reporter / genetics* Genes, Reporter / physiology* Green Fluorescent Proteins / metabolism* Integrases / genetics* Integrases / metabolism* Islets of Langerhans / metabolism Luminescent Proteins Male Mice Mice, Inbred C57BL Mice, Transgenic Recombination, Genetic Ubiquitination
Resource
DNA material pROSA26CAG-GFP-tdsRed (RDB19441)