TMC2-AcGFP mice express fluorescent protein in inner ear sensory hair cells.
Dr. Andrew J. Griffith, NIDCD/NIH
Genetic Background | Generation |
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Allele Type |
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Transgenic (Reporter, Inserted expressed sequence) |
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.
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.
Expressed Gene | GFP, Green Fluorescent Protein, |
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Expressed Gene | Tmc2, transmembrane channel-like gene family 2, mouse, laboratory |
Site of Expression | Inner ear sensory hair cells. |
Allele Name | transgene insertion 3, Andrew Griffith |
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Allele Type | Transgenic (Reporter, Inserted expressed sequence) |
Allele Synonym(s) | Tg(RP24-293G8/Tmc2/AcGFP)3Ajg; Tmc2-AcGFP |
Gene Symbol and Name | Tg(Tmc2/AcGFP)3Ajg, transgene insertion 3, Andrew Griffith |
Gene Synonym(s) | |
Promoter | Tmc2, transmembrane channel-like gene family 2, mouse, laboratory |
Expressed Gene | GFP, Green Fluorescent Protein, |
Expressed Gene | Tmc2, transmembrane channel-like gene family 2, mouse, laboratory |
Site of Expression | Inner ear sensory hair cells. |
Strain of Origin | (C57BL/6 x SJL)F2 |
Chromosome | UN |
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. |
Hemiygotes and homozygotes are viable and fertile.
When using the TMC2-AcGFP mouse strain in a publication, please cite the originating article(s) and include JAX stock #028517 in your Materials and Methods section.
Facility Barrier Level Descriptions
Service/Product | Description | Price |
---|---|---|
Hemizygous or Non carrier for Tg(Tmc2/AcGFP)3Ajg |
Frozen Mouse Embryo | B6.Cg-Tg(Tmc2/AcGFP)3Ajg/J Frozen Embryo | $2595.00 |
Frozen Mouse Embryo | B6.Cg-Tg(Tmc2/AcGFP)3Ajg/J Frozen Embryo | $2595.00 |
Frozen Mouse Embryo | B6.Cg-Tg(Tmc2/AcGFP)3Ajg/J Frozen Embryo | $3373.50 |
Frozen Mouse Embryo | B6.Cg-Tg(Tmc2/AcGFP)3Ajg/J Frozen Embryo | $3373.50 |
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