B6N(Cg)-Vim^tm1Ggen/J

Stock number: 039691
Product name: Vim^{FLEX(SA-eGFP-T2A-HSV-TK-WPRE-pA)}
Strain synonyms: Vim^TK, Vim-GFP-TK
Research areas: Cancer Research, Cell Biology Research, Research Tools

Description

Vim^{FLEX(SA-eGFP-T2A-HSV-TK-WPRE-pA)} mice have inward facing lox2272 sites and loxP sites flanking exons 2-4 of the Vim gene and an inverted cassette containing a polyadenylation sequence, an HSV TK (herpes simplex 1 virus thymidine kinase), a T2A sequence, and an EGFP (enhanced green fluorescent protein). These Vim^{FLEX(SA-eGFP-T2A-HSV-TK-WPRE-pA)} mice express endogenous wildtype VIM prior to Cre-mediated recombination. Following Cre-mediated recombination, exons 2-4 are removed and the TK-EGFP knock-in cassette is inverted into the correct orientation for expression from the Vim promoter. Expression of TK allows for ganciclovir-inducible ablation of Vim-expressing cells. This strain may be useful for studying the role of VIM in cell migration, cell shape and plasticity, and Epithelial-to-Mesenchymal Transition (EMT).

The donating laboratory has also made a Cre-inducible Vim^{FLEX(SA-H2B-eGFP-T2A-FlpO-WPRE-pA)} mouse line available as Stock No. 039724.

Detailed description

Vim^{FLEX(SA-eGFP-T2A-HSV-TK-WPRE-pA)} mice have a lox2272 site upstream of exon 2 of the Vimentin (Vim) gene and an inverted cassette flanked with inward facing loxP sites downstream of exon 4. The cassette contains a polyadenylation sequence, an HSV TK (herpes simplex 1 virus thymidine kinase), a T2A sequence, an EGFP (enhanced green fluorescent protein), a splice acceptor site and a second lox2272 site in opposite orientation to the first, followed by an Frt-flanked neomycin cassette and a second inward facing loxP site. Vimentin is a structural protein expressed by mesenchymal cells and plays a role in cell migration, cell shape and plasticity, and Epithelial-to-Mesenchymal Transition (EMT). These Vim^{FLEX(SA-eGFP-T2A-HSV-TK-WPRE-pA)} mice express endogenous wildtype VIM prior to Cre-mediated recombination. Following Cre-mediated recombination, exons 2-4 are removed and the TK-EGFP knock-in cassette is inverted into the correct orientation for expression from the Vim promoter. Expression of TK allows for ganciclovir-inducible ablation of Vim-expressing cells.

Homozygous Vim^{FLEX(SA-eGFP-T2A-HSV-TK-WPRE-pA)} mice are viable and fertile with no overt phenotype. Following Cre-mediated recombination, EGFP is detectable by immunofluorescence and by fluorescence-activated cell sorting (FACS).

The donating laboratory has also made a Cre-inducible Vim^{FLEX(SA-H2B-eGFP-T2A-FlpO-WPRE-pA)} mouse line available as Stock No. 039724.

Development

The Vim^{FLEX(SA-eGFP-T2A-HSV-TK-WPRE-pA)} targeting vector was designed with (from 5' to 3') an inward facing lox2272 site upstream of exon 2 of the vimentin (Vim) gene, followed by exons 3 and 4, and an inverted cassette flanked with inward facing loxP sites. The inverted floxed cassette contains a polyadenylation sequence, an HSV TK (herpes simplex 1 virus thymidine kinase), a T2A sequence, an EGFP (enhanced green fluorescent protein), a splice acceptor site and a second lox2272 site in opposite orientation to the first, followed by an FRT-flanked neomycin cassette and a second inward facing loxP site all downstream of exon 4. This construct was electroporated into C57BL/6N-derived embryonic stem (ES) cells. Correctly targeted ES cells were injected into blastocysts, and resulting chimeric mice were bred to Flp-deleter mice to remove the Neo cassette, and then to C57BL/6N animals for 10 generations by the 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/6NJ oocytes (Stock No. 005304).

Allele

Symbol: Vim^tm1Ggen
Name: targeted mutation 1, Giannicola Genovese
MGI accession ID: MGI:7718775
Generation method: Targeted
Attributes: Conditional ready (e.g. floxed); Reporter
Synonyms: Vim^{FLEX(SA-eGFP-T2A-HSV-TK-WPRE-pA)}; Vim^{FLEX(SA-TK-T2AeGFP-pA)}; Vim^TK
Marker symbol: Vim
Marker name: vimentin
Chromosome: 2
Strain of origin: C57BL/6N
Molecular note: The targeting vector was designed with (from 5'to 3') an inward facing lox2272 site upstream of exon 2 of the vimentin(Vim) gene, followed by exons 3 and 4, and an inverted cassette flanked with inward facing loxP sites. The inverted floxed cassette contains a polyadenylation sequence, an HSV TK (herpes simplex 1 virus thymidine kinase), a T2A sequence, an EGFP (enhanced green fluorescent protein), a splice acceptor site and a second lox2272 site in opposite orientation to the first, followed by an FRT-flanked neomycin cassette and a second inward facing loxP site all downstream of exon 4.