STOCK Tg(tetO-hop/EGFP,-COP4/mCherry)6Kftnk/J

Stock number: 017906
Research areas: Neurobiology Research, Research Tools

Description

BTBR6 transgenic mice have a bidirectional tetracycline-responsive promoter (TRE; tetO) driving expression of two individual fusion genes; the halorhodopsin/EGFP fusion gene (HaloR-EGFP) and channelrhodopsin-2/mCherry fusion gene (ChR2-mCherry). When bred to other mice expressing tTA or reverse tetracycline-controlled transactivator protein (rtTA), simultaneous HaloR-EGFP and ChR2-mCherry expression in the resulting double mutant offspring can be regulated by tetracycline or its analog doxycycline (dox). These mice may be useful for optogenetic studies visualizing neural circuits.

Detailed description

BTBR6 transgenic mice have a bidirectional tetracycline-responsive promoter (TRE; tetO) driving expression of two individual fusion genes; the halorhodopsin/EGFP fusion gene (HaloR-EGFP) and channelrhodopsin-2/mCherry fusion gene (ChR2-mCherry). When bred to other mice expressing tTA or reverse tetracycline-controlled transactivator protein (rtTA), simultaneous HaloR-EGFP and ChR2-mCherry expression in the resulting double mutant offspring can be regulated by tetracycline or its analog doxycycline (dox). The specificity of the tTA strain chosen to generate the bi-transgenic line confers inducible conditional expression of the transgene, and via photostimulation expression of Channelrhodopsin-2, in defined cell populations.
When crossed with a Camk2a-tTA strain (see for example, Stock No. 003010), double transgenic mice show specific expression of EGFP and mCherry in the dorsal striatum and projections into the globus pallidus and substantia nigra, allowing for visualization of neuronal excitability and neural circuits.

Development

A transgenic construct containing a SV40 polyadenylation site sequence (reverse orientation) followed by the halorhodopsin-enhanced green fluorescent protein fusion gene cDNA, and the Channelrhodopsin-2 (Chlamydomonas reinhardtii)/mCherry, Red Fluorescent Protein fusion gene cDNA followed by a SV40 polyadenylation site sequence, under the control of the a bidirectional tetracycline-responsive promoter (TRE; tetO) was injected into fertilized CBA;B6 mouse eggs. Founder line 6 was subsequently established. The mice were then crossed to mice on the B6;129 background for several generations. Upon arrival at The Jackson Laboratory, the mice were crossed to C57BL/6J (Stock No. 000664) at least once to establish the colony.

Allele

Symbol: Tg(tetO-hop/EGFP,-COP4/mCherry)6Kftnk
Name: transgene insertion 6, Kenji Tanaka
MGI accession ID: MGI:5439158
Generation method: Transgenic
Attributes: Reporter; Inducible
Marker symbol: Tg(tetO-hop/EGFP,-COP4/mCherry)6Kftnk
Marker name: transgene insertion 6, Kenji Tanaka
Chromosome: UN
Strain of origin: CBA x C57BL/6
Expressed genes: HOP,Halorhodopsin,Natronomonas; GFP,Green Fluorescent Protein,; COP4,Channelrhodopsin,Chlamydomonas; RFP,Red Fluorescent Protein,coral
Site of expression: When mated to a mutant strain expressing tetracycline-controlled transactivator protein (tTA), expression of HOP/EGFP,COP4/mCherry may be regulated with tetracycline or its analog doxycycline (dox) in the double mutant offspring. The specificity of the tTA strain chosen to generate the bi-transgenic line confers inducible conditional expression of the transgene, and via photostimulation expression of Channelrhodopsin-2, in defined cell populations. For example, when crossed with a Camk2a-tTA strain, double transgenic mice show specific expression of EGFP and mCherry in the dorsal striatum and projections into the globus pallidus and substantia nigra.
Molecular note: A bidirectional tetracycline responsive element drives inducible expression of a halorhodopsin/EGFP fusion and a channelrhodopsin-2/m-Cherry fusion. Line 6 was the only line to exhibit expression.