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

Summary

• Symbol: Igs7^{tm136.1(tetO-COP3*/COP4*/EYFP)Hze}
• Name: intergenic site 7; targeted mutation 136.1, Hongkui Zeng
• MGI ID: MGI:5903984
• Synonyms: Ai136D, Ai136(TITL-ReaChR-YFP)-D
• Gene: Igs7 Location: unknown Genetic Position: Chr9, Syntenic
• Alliance: Igs7^{tm136.1(tetO-COP3*/COP4*/EYFP)Hze} page

Mutation origin

• Germline Transmission: Earliest citation of germline transmission: J:260362
• Parent Cell Line: G4 (ES Cell)
• Strain of Origin: (129S6/SvEvTac x C57BL/6NCrl)F1

Mutation description

• Allele Type: Targeted (Conditional ready, Inducible, Reporter)
• Inducer: doxycycline/tetracycline
• Mutation: Insertion
• Mutation details: 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 carterichannelrhodopsin 1 [cop3]), transmembrane domain F of VChR2 (V. carterichannelrhodopsin 2 [cop4]) and the ChEF/ChIEF variant N-terminal extracellular sequences (derived fromChlamydomonas reinhardtiichannelrhodopsin 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]). (J:101977, J:260362)

Find Mice (IMSR)

• Mouse strains and cell lines available from the International Mouse Strain Resource (IMSR)
• Carrying this Mutation: Mouse Strains: 1 strain available Cell Lines: 0 lines available
• Carrying any Igs7 Mutation: 70 strains or lines available

References

• Original: J:260362 Daigle TL, et al., A suite of transgenic driver and reporter mouse lines with enhanced brain cell type targeting and functionality. Cell. 2018 Jul 12;174(2):465-480.e22
• All: 2 reference(s)