Igs7^{tm90.1(tetO-COP4*/EGFP)Hze}
Targeted Allele Detail

Summary

• Symbol: Igs7^{tm90.1(tetO-COP4*/EGFP)Hze}
• Name: intergenic site 7; targeted mutation 90.1, Hongkui Zeng
• MGI ID: MGI:5558069
• Synonyms: Ai90D, Ai90deltaHygro, Ai90(TITL-Chronos)-D, Ai90(TITL-ShChR)-D
• Gene: Igs7 Location: unknown Genetic Position: Chr9, Syntenic
• Alliance: Igs7^{tm90.1(tetO-COP4*/EGFP)Hze} page

Mutation origin

• Germline Transmission: Earliest citation of germline transmission: J:101977
• 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 targeting vector was designed with (from 5' to 3') an FRT3 site, two tandem copies of the chicken beta-globin HS4 insulator element (to reduce reporter gene expression in absence of transactivator protein), a modified Tet response element (TRE or tetO), a loxP-flanked STOP cassette (stop codons in all three reading frames linked to synthetic pA-hGHpA-PGKpA), the COP4*/EGFP coding sequence, a WPRE sequence (to enhance mRNA transcript stability), a bovine growth hormone polyA signal, two tandem copies of the chicken beta-globin HS4 insulator element, an AttB site, a PGK-5' hygro cassette, an RNA splice donor and an FRT5 site. The COP4*/EGFP fusion protein is composed of a mammalian codon-optimized Stigeoclonium helveticum-derived channelrhodopsin ShChR (Chronos) fused in-frame via a 18 nucleotide linker to the N-terminus of enhanced green fluorescent protein (EGFP). Chronos is a blue-to-green light-driven (~470-530 nm) cation channel that depolarizes the cell and causes action potentials. Chronos has high light sensitivity and faster kinetics than previous channelrhodopsins. Embryonic stem cells, previously targeted with FRT3-AttB-PGK-neoR-polyA::FRT5::splice acceptor::3' hygro cassette::SV40 polyA: AttP in the same locus, were re-transfected with the targeting vector and Flp recombinase vector. Mutant mice were bred with Gt(ROSA)26Sor^{tm3(phiC31*)Sor} mice to remove to remove the AttB/AttP-flanked PGK-hygro-SV40pA cassette and replace it with the recombined AttB/AttP site (AttL). The Ai90D allele is therefore TIGRE-frt3-Ins-TRE-LSL-Chronos/EGFP-WPRE-bGHpA-Ins-AttL. (J:101977, J:219930)

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: 66 strains or lines available

References

• Original: J:101977 The Jackson Laboratory, Information obtained from The Jackson Laboratory, Bar Harbor, ME. Unpublished. 2005-2017
• All: 4 reference(s)