Igs7tm143.1(tetO-RCaMP1.07)Hze
Targeted Allele Detail
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Symbol: |
Igs7tm143.1(tetO-RCaMP1.07)Hze |
Name: |
intergenic site 7; targeted mutation 143.1, Hongkui Zeng |
MGI ID: |
MGI:5903975 |
Synonyms: |
Ai143D, TITL-R-CaMP1.07-D |
Gene: |
Igs7 Location: unknown Genetic Position: Chr9, Syntenic
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Alliance: |
Igs7tm143.1(tetO-RCaMP1.07)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 replacement vector is designed with (from 5' to 3') an FRT3 site, two copies of chicken beta-globin HS4 insulator element, a modified Tet response element/promoter, a floxed-STOP cassette, the R-CaMP1.07 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-R-CaMP1.07 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).
R-CaMP1.07 is a red calcium-sensing molecule that functions at cellular calcium levels and is a modified version of R-CaMP1.01 (which was created by insertion of random mutations into R-GECO1). R-CaMP1.07 is composed (from N-term to C-term) of a 35 aa polyHis plasmid leader sequence, a 13 aa chicken smooth muscle myosin light chain kinase, a circularly permutated red fluorescent protein (mApple; aa 146-231 followed by aa 1-145), a rat calmodulin DNA fragment, a 15 aa linker and a self-cleaving peptide (F2A).
(J:241918)
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Original: |
J:241918 Bethge P, et al., An R-CaMP1.07 reporter mouse for cell-type-specific expression of a sensitive red fluorescent calcium indicator. PLoS One. 2017;12(6):e0179460 |
All: |
4 reference(s) |
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