Igs7tm93.1(tetO-GCaMP6f)Hze
Targeted Allele Detail
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Symbol: |
Igs7tm93.1(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
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Alliance: |
Igs7tm93.1(tetO-GCaMP6f)Hze page
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Allele Type: |
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Targeted (Conditional ready, Inducible, Reporter) |
Inducer: |
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doxycycline/tetracycline |
Mutation: |
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Insertion
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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)26Sortm3(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)
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View phenotypes and curated references for all genotypes (concatenated display).
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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: |
48 reference(s) |
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