C57BL/6-Tg(CAG-tdTomato,Actb/EGFP)1Erv/J

Stock number: 029889
Product name: FLEx-actin-GFP
Research areas: Neurobiology Research, Research Tools

Description

FLEx β-actin-GFP mice have widespread tdTomato expression. Following Cre-mediated recombination, tdTomato is removed and mice conditionally express β-actin-GFP. This strain may be useful for monitoring actin dynamics.

Detailed description

FLEx β-actin-GFP mice were created using a cre-dependent one-way genetic switch (FLEx) system. Hemizygotes mice carrying this conditional allele are viable and fertile. Two sets of incompatible inward facing recombination sites (lox2372 and lox511) flank a tdTomato sequence and an inverted β-actin-GFP. Prior to Cre recombination mice have widespread expression of tdTomato. Because this construct is driven by a ubiquitous CAG promoter, β-actin-GFP expression is determined by which tissue(s) express Cre recombinase. When bred to mice that express Cre recombinase, the resulting offspring will have the lox2372 or lox511 sites recombined, resulting in the excision of the intervening tdTomato sequence, and the expression of the β-actin-GFP sequence, in a sense orientation.

Development

The FLEx β-actin-GFP targeting vector was designed with (from 5’ to 3’) a, CMV enhancer chicken beta-actin (CAG) promoter, a lox2372 site, a tdTomato red fluorescent protein sequence, and a lox511 site in forward orientation. This is followed by Actb (actin, beta) and EGFP (enhanced green fluorescent protein), a lox2372 site, and a lox511 site all in reverse orientation, and a polyadenylation signal in forward orientation. This entire construct was injected into C57BL/6 oocytes, and founder mice were bred to C57BL/6 mice to establish a colony of FLEx-actin-GFP mice. Mice were backcrossed for 10 generations to C57BL/6J mice. 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 oocytes (Stock No. 000664).

Allele

Symbol: Tg(CAG-tdTomato,Actb/EGFP)1Erv
Name: transgene insertion 1, James Ervasti
MGI accession ID: MGI:5810463
Generation method: Transgenic
Attributes: Conditional ready (e.g. floxed); Reporter
Synonyms: beta-actin-GFP
Marker symbol: Tg(CAG-tdTomato,Actb/EGFP)1Erv
Marker name: transgene insertion 1, James Ervasti
Chromosome: UN
Strain of origin: C57BL/6
Expressed genes: RFP,Red Fluorescent Protein,coral
Site of expression: Prior to Cre recombination, mice have widespread expression of tdTomato. When these mutant mice are bred to mice that express Cre recombinase, the resulting offspring will express GFP in the cre-expressing tissues
Molecular note: The targeting vector is designed with (from 5' to 3') a, CMV enhancer chicken beta-actin (CAG) promoter, a lox2372 site, a tdTomato red fluorescent protein sequence, and a lox511 site in forward orientation. This is followed by mouse Actb (actin, beta) and EGFP (enhanced green fluorescent protein), a lox2372 site, and a lox511 site all in reverse orientation, and a polyadenylation signal in forward orientation.