STOCK Aldh1a3^{tm1(cre/ERT2)Dac}/J

Stock number: 039535
Product name: Aldh1a3-Cre^ert
Research areas: Cancer Research, Diabetes and Obesity Research, Research Tools

Description

The Aldh1a3-Cre^ert knock-in mice have a tamoxifen-activable Cre (Cre^ert) knocked into the endogenous aldehyde dehydrogenase family 1, subfamily A3 (Aldh1A3) gene locus. The line allows for tamoxifen-inducible Cre recombinase activities in cells expressing Aldh1a3. The line is useful for lineage tracing and the study of Aldh1a3 role in dedifferentiating pancreatic β-cells.

Detailed description

The Aldh1a3-Cre^ert knock-in mice have a P2A linker and a tamoxifen-activable Cre (Cre^ert) inserted just downstream of the endogenous aldehyde dehydrogenase family 1, subfamily A3 (Aldh1A3) gene locus. The line allows for tamoxifen-inducible Cre recombinase activities in cells expressing Aldh1a3. ALDH1A3 (also known as RALDH3) is a marker of β-cell dedifferentiation in murine and human type 2 diabetes patients. ALDH1A3-positive β-cells show reduced insulin secretion compared to ALDH1A3-negative β-cells. The donating laboratory reported crossing to the mice to R26-stop-EYFP line mice (Stock No. 006148) and Lepr^db (Stock No. 000697) labeled over 20% of pancreatic islet cells after tamoxifen administration. The homozygous mice are viable and fertile. These Aldh3a1-Cre^ert knock-in mice may be useful for lineage tracing and the study of Aldh1a3 role in dedifferentiating pancreatic β-cells and diabetes research.

Development

A targeting construct was designed to insert the coding sequence for tamoxifen-inducible Cre recombinase fused with ERT2 and frt-flanked neo selection cassette downstream of the stop sequence of the Aldh1a3 locus with a P2A self-cleaving peptide separating the Aldh1a3 and cre/ERT2 coding sequences. The targeting construct was injected into 129B6 hybrid-derived ES cells. The ES clones with correct integration were confirmed by PCR and Sanger sequencing and introduced into recipient mice blastocysts. The chimeric female mice were bred with ACTB-Flpe male mice to remove the frt-flanked neo cassette. Correctly targeted mice were further bred with C57BL/6 mice 3-4 generations to generate the Aldh1a3-Cre^ert mouse line and maintained in C57BL/6 genetic background by donating laboratory. Upon arrival at The Jackson Laboratory, sperm was cryopreserved. To establish our live colony, an aliquot of frozen sperm was used to fertilize C57BL/6J (Stock No. 000664) oocytes.