B6.Cg-Tg(Tmc2/AcGFP)3Ajg/J

Stock number: 028517
Product name: TMC2-AcGFP
Research areas: Research Tools

Description

TMC2-AcGFP mice express fluorescent protein in inner ear sensory hair cells.

Detailed description

Transmembrane channel-like gene family 2 (Tmc2) is essential for mechanoelectrical transduction (MET) in inner ear sensory hair cells.

These TMC2-AcGFP BAC transgenic mice express the mouse Tmc2 gene and an AcGFP fluorescent reporter under the control of the endogenous Tmc2 promoter. Expression of the transgene was confirmed in the presumed site of mechanolelectrical transduction at stereocilia tips. Confocal images of inner hair cell stereocilia from TMC2-AcGFP transgenic mice demonstrate the protein localizes at the tips of vestibular hair cell stereocilia at postnatal day 2 (P2). By P3, fluorescent puncta are abundant and localized along the length of stereocilia as well as at the tips. At P7, TMC2-AcGFP puncta are less evident in outer hair cell (OHC) stereocilia, but present at stereocilia tips of inner hair cells (IHCs). By P10, puncta disappear from IHC stereocilia.

Development

The Tmc2 stop codon (c. 2665-2667) of mouse bacterial artificial chromosome (BAC) RP24-293G8 was replaced with cDNA encoding AcGFP (a mammalian-optimized, monomeric fluorphore derived from Aequorea coerulescens). The modified BAC, also carrying the Snrpb, Nop56, and Idh3b genes, was purified and injected into (C57BL/6 x SJL) F2 mouse embryos. Founder mice carrying the transgene were intercrossed with other targeted and transgenic alleles (see Stock Nos. 028392, 019146, and/or 019147). The TMC2-AcGFP transgene was subsequently bred away from the other mutations. Line 3, carrying 3 copies of the transgene was backcrossed from a mixed background (C57BL/6, SJL, 129) to C57BL/6 for more than 5 generations by the donating lab.

Allele

Symbol: Tg(Tmc2/AcGFP)3Ajg
Name: transgene insertion 3, Andrew Griffith
MGI accession ID: MGI:5560295
Generation method: Transgenic
Attributes: Reporter; Inserted expressed sequence
Marker symbol: Tg(Tmc2/AcGFP)3Ajg
Marker name: transgene insertion 3, Andrew Griffith
Chromosome: UN
Strain of origin: (C57BL/6 x SJL)F2
Expressed genes: GFP,Green Fluorescent Protein,; Tmc2,transmembrane channel-like gene family 2,mouse, laboratory
Site of expression: Inner ear sensory hair cells.
Molecular note: The transgene was created by targeted insertion of Aequorea coerulescens green fluorescent protein (AcGFP) coding sequence into the bacterial artificial chromosome (BAC) RP24-293G8. The AcGFP sequence integrated at the stop codon of Tmc2. The integration site was not investigated. The C-terminal fusion is expressed in the sensory hair cells of the inner ear. The modified BAC also carries the Snrpb, Nop56, and Idh3b genes. Line 3 carries 3 copies of the transgene.