B6.Cg-Igs7^{tm136.1(tetO-COP3*/COP4*/EYFP)Hze}/J

Igs7^{tm136.1(tetO-COP3*/COP4*/EYFP)Hze}

targeted mutation 136.1

Conditional ready (e.g. floxed); Reporter; Inducible

Igs7

intergenic site 7

9

YFP, Yellow Fluorescent Protein, jellyfish

5903984

When bred with other mice expressing Cre recombinase, tetracycline-controlled transactivator protein (tTA) and/or reverse tetracycline-controlled transactivator protein (rtTA), ReaChR expression in cells/tissues where the expression patterns of the individual promoters driving Cre and tTA/rtTA overlap can be regulated with tetracycline or its analog doxycycline (dox).

(129S6/SvEvTac x C57BL/6NCrl)F1

The replacement vector was designed with (from 5' to 3') an FRT3 site, two copies of chicken beta-globin HS4 insulator element, a modified Tet response element/promoter (TRE/tetO), a floxed-STOP cassette (stop codons in all three reading frames linked to synthetic pA-hGHpA-PGKpA), the ReaChR/EYFP sequence (details below), a WPRE, a BGH polyA, two copies of chicken beta-globin HS4 insulator element, an AttB site, a PGK-5'hygro cassette, an RNA splice donor and a FRT5 site.Embryonic stem cells previously targeted with FRT3::AttB::PGK-neoR-polyA::FRT5::splice acceptor::3'hygro cassette::SV40 polyA:AttP in TIGRE, were re-transfected with the pTRE-LSL-ReaChR/EYFP replacement vector and Flp recombinase vector for recombinase-mediated cassette exchange (RCME).PhiC31-mediated recombination removed the AttB/AttP-flanked sequence (PGK-Neo-polyA::frt5::RNA splice acceptor::3'hygro-polyA) and replaced it with the recombined AttB/AttP site (AttL). The ReaChR/EYFP fusion protein has the red-activatable channelrhodopsin variant ReaChR fused in-frame at its C-terminus (via a G-S-G linker) to an enhanced yellow fluorescent protein (EYFP). ReaChR is a chimeric channelrhodopsin composed of transmembrane domains A-E and G of VChR1 (Volvox carteri channelrhodopsin 1 [cop3]), transmembrane domain F of VChR2 (V. carteri channelrhodopsin 2 [cop4]) and the ChEF/ChIEF variant N-terminal extracellular sequences (derived from Chlamydomonas reinhardtii channelrhodopsin 1 [cop3]). The ChEF/ChIEF, VChR1 and VChR2 sequences are mammalian-codon optimized, and transmembrane domain C harbors two point mutations encoding I170V (to accelerate rate of channel closure while retaining reduced inactivation) and L171I (to enhance activation by red light [~590 nm]).

C57BL/6J

The donating investigator reports that heterozygous and homozygous mice are viable and fertile with no reported gross physical or behavioral abnormalities. When maintaining a live colony, heterozygous mice may be bred together, to wildtype mice from the colony or to C57BL/6J inbred mice. Alternatively, homozygous mice may be bred together.

Homozygote x Homozygote