Igs7^{tm169.1(tetO-GFP*/TPTE2*,CAG-tTA2)Hze}
Targeted Allele Detail

Summary

• Symbol: Igs7^{tm169.1(tetO-GFP*/TPTE2*,CAG-tTA2)Hze}
• Name: intergenic site 7; targeted mutation 169.1, Hongkui Zeng
• MGI ID: MGI:6151064
• Synonyms: Ai169, Ai169(TIT2L-ASAP2s-ICL-tTA2)
• Gene: Igs7 Location: unknown Genetic Position: Chr9, Syntenic
• Alliance: Igs7^{tm169.1(tetO-GFP*/TPTE2*,CAG-tTA2)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, Transactivator)
• Inducer: doxycycline/tetracycline
• Mutation: Insertion
• Mutation details: The vector is designed with (from 5' to 3') an frt3 site, two copies of chicken beta-globin HS4 insulator element (to reduce reporter gene expression in absence of transactivators), a Tet response element/promoter (TRE2; details below), a loxP-flanked STOP cassette (stop codons in all three reading frames linked to synthetic pA-hGHpA-PGKpA), a green fluorescent voltage indicator protein ASAP2s sequence (details below), a WPRE (to enhance the mRNA transcript stability), a BGH polyA, two copies of chicken beta-globin HS4 insulator element, a CMV-IE enhancer/chicken beta-actin/rabbit beta-globin hybrid promoter (CAG; from pCAGGS), a lox2272-flanked STOP cassette (stop codons in all three reading frames linked to synthetic pA-hGHpA-TKpA), a synthetic modified tetracycline-regulated transactivator gene (tTA2^S), a WPRE, a BGH polyA, an AttB site, a PGK-5'hygro cassette, an RNA splice donor and a FRT5 site. The TRE2 promoter used here is Tet-responsive P>hCMV*-1<; containing the Tet response element (seven copies of the 19 bp tet operator sequence [tetO]) just upstream of a minimal cytomegalovirus promoter (P>min CMV<), which lacks the enhancer that is part of the complete CMV promoter. Consequently, P>hCMV*-1< is silent in the absence of tTA or rtTA binding to tetO. GFP*/TPTE2* has VSD (voltage sensor domwin) sequences (GgVSD) from the Gallus gallus voltage-sensitive phosphatase TPTE2, including the R154Q mutation for desired voltage response. A circularly permuted superfolder GFP 1-10 OPT variant (cpsfGFP-OPT) is inserted into the GgVSD extracellular loop between the third and fourth transmembrane segments (S3-S4). The GgVSD includes the R154Q mutation for desired voltage response (equivalent to R153Q in original descriptions). The GFP 1-10 OPT includes the substitutions from folding reporter GFP (F99S, M153T, V163A, F64L, S65T) and superfolder GFP (S30R, Y145F, I171V, A206V), and also has seven additional substitutions (N39I, T105K, E111V, I128T, K166T, I167V, S205T). Together, these mutations result in improved brightness and dynamic range, while maintaining efficient expression at the membrane. PhiC31-mediated recombination removed the AttB/AttP-flanked sequence and replaced it with the recombined AttB/AttP site (AttL). (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)