Gt(ROSA)26Sor^{tm27.1(CAG-COP4*H134R/tdTomato)Hze}
Targeted Allele Detail

Summary

• Symbol: Gt(ROSA)26Sor^{tm27.1(CAG-COP4*H134R/tdTomato)Hze}
• Name: gene trap ROSA 26, Philippe Soriano; targeted mutation 27.1, Hongkui Zeng
• MGI ID: MGI:4834650
• Synonyms: Ai27, Ai27DeltaNeo, Ai27D (Rosa-CAG-LSL-hChR2(H134R)-tdTomato-WPRE), ChR2(H134R)-tdTomato, Rosa26-lsl-ChR2-tdTomato
• Gene: Gt(ROSA)26Sor Location: Chr6:113044389-113054205 bp, - strand Genetic Position: Chr6, 52.73 cM
• Alliance: Gt(ROSA)26Sor^{tm27.1(CAG-COP4*H134R/tdTomato)Hze} page

Mutation origin

• Germline Transmission: Earliest citation of germline transmission: J:182204
• Parent Cell Line: G4 (ES Cell)
• Strain of Origin: (129S6/SvEvTac x C57BL/6NCrl)F1

Mutation description

• Allele Type: Targeted (Conditional ready, Reporter)
• Mutation: Insertion
• Mutation details: The Rosa-CAG-LSL-hChR2(H134R)-tdTomato-WPRE targeting vector was designed with (from 5' to 3') a CMV-IE enhancer/chicken beta-actin/rabbit beta-globin hybrid promoter (CAG), an FRT site, a loxP-flanked STOP cassette (with stop codons in all 3 reading frames and a triple polyA signal), a mammalianized ChR2(H134R)-tdTomato fusion gene, a woodchuck hepatitis virus post-transcriptional regulatory element (WPRE; to enhance the mRNA transcript stability), a BGH polyA signal, and an attB/attP-flanked PGK-FRT-Neo-polyA cassette. To create the mammalianized ChR2(H134R)-tdTomato fusion gene, a cDNA sequence encoding the first 315 amino acids of channelrhodopsin-2 (derived from the green alga Chlamydomonas reinhardtii) was modified to harbor condons optimized for mammalian expression and a gain-of-function H134R substitution (CAC to CGC) designed to cause larger stationary photocurrents. This mhChR2 sequence (also called hChR2(H134R) or hChR2-H134R) was fused in-frame to the amino terminus of a tdTomato sequence (a non-oligomerizing DsRed fluorescent protein variant with a 12 residue linker fusing two copies of the protein (tandem dimer)), resulting in the final hChR2(H134R)-tdTomato fusion protein sequence. The entire Rosa-CAG-LSL-hChR2(H134R)-tdTomato-WPRE targeting vector was inserted between exons 1 and 2 of the locus. PhiC31 mediated recombination removed the PGK-FRT-Neo-polyA cassette. (J:182204)

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 Gt(ROSA)26Sor Mutation: 1040 strains or lines available

References

• Original: J:182204 Madisen L, et al., A toolbox of Cre-dependent optogenetic transgenic mice for light-induced activation and silencing. Nat Neurosci. 2012 Mar 25;15(5):793-802
• All: 111 reference(s)