Igs7^{tm93.1(tetO-GCaMP6f)Hze}
Targeted Allele Detail

Summary

• Symbol: Igs7^{tm93.1(tetO-GCaMP6f)Hze}
• Name: intergenic site 7; targeted mutation 93, Hongkui Zeng
• MGI ID: MGI:5558086
• Synonyms: Ai93, Ai93D, Ai93(TITL-GCaMP6f)
• Gene: Igs7 Location: unknown Genetic Position: Chr9, Syntenic
• Alliance: Igs7^{tm93.1(tetO-GCaMP6f)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 GCaMP6 fast variant calcium indicator (GCaMP6f) coding sequence, a WPRE sequence (to enhance the 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 GCaMP6 fast variant calcium indicator (GCaMP6f) is a detector of single neuronal action potentials with fast response kinetics, and is an improved version of GCaMP5G. GCaMP6f has a chicken smooth muscle M13 fragment of myosin light chain kinase, a circularly permutated EGFP, and a rat calmodulin DNA fragment mutation for accelerated kinetics. Embryonic stem cells, previously targeted with FRT5-AttB-Hygro2 cassette-AttP-SV40pA-splice acceptor-PGKpA-FRT3 in the same locus, were re-transfected with the targeting vector and Flp recombinase vector. Genotyping/sequencing revealed that Ai93 had two tandem copies of the pTRE-LSL-GCaMP6f replacement vector inserted during RCME. Chimeric mice were bred with PhiC31 Gt(ROSA)26Sor^{tm3(phiC31*)Sor} mice to remove the AttB/AttP-flanked region. The Ai93D allele is therefore TIGRE-frt3-Ins-TRE-LSL-GCaMP6f-WPRE-bGHpA-Ins-AttB-PGK-5'hygro-SD-frt5-frt3-Ins-TRE-LSL-GCaMP6f-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: 3 strains available Cell Lines: 0 lines available
• Carrying any Igs7 Mutation: 63 strains or lines available

References

• Original: J:219930 Madisen L, et al., Transgenic mice for intersectional targeting of neural sensors and effectors with high specificity and performance. Neuron. 2015 Mar 4;85(5):942-58
• All: 38 reference(s)