B6;129-Gt(ROSA)26Sortm(cMYC,-EGFP)Mro/H
| Status | Available to order |
| EMMA ID | EM:15260 |
| International strain name | B6;129-Gt(ROSA)26Sortm(cMYC,-EGFP)Mro/H |
| Alternative name | STOPFloxc-MYC |
| Strain type | Targeted Mutant Strains : Conditional mutation |
| Allele/Transgene symbol | Gt(ROSA)26Sortm(cMYC,-EGFP)Mro |
| Gene/Transgene symbol | Gt(ROSA)26Sor |
Information from provider
| Provider | Silvia Marino |
| Provider affiliation | Centre for Genomic and Child Health, Blizard |
| Genetic information | Conditional gain of function expression of c-MYC. The Gateway Entry system was chosen to guarantee controlled and efficient monosite insertion of the c-MYC construct into the ubiquitously expressed Gt(ROSA)26Sor locus. The toxin-encoding ccdB gene was excised from the pENTR1A vector (Invitrogen) by BamH1/Xho1 restriction digest and a 1.5 kb cDNA construct of human c-MYC was inserted into these sites of the pENTR1A vector. BamH1/Xho1 restriction digest showed correct insertion of the construct. Correct orientation of the insert was confirmed by DNA sequencing. The c-MYC construct was subsequently inserted into a targeting vector via in vitro recombination. Since the c-MYC construct in the pENTR1A vector is flanked by specific lambda phage integrase recognition sites (attL), the c-MYC construct could be efficiently transferred to a targeting vector carrying the corresponding heterotypic sites (attR). The targeting vector contains a 5′ homology region to the mouse Gt(ROSA)26Sor genomic locus, a splice acceptor (SA) site, a PGK-neo-3x pA stop cassette flanked by loxP-sites (LSL), the c-MYC construct, an IRES-eGFP reporter gene, a 3′ homology region to the mouse ROSA26 genomic sequence and a Diphteria Toxin A (DTA) selection cassette. It was electroporated into G4 F1 hybrid ES cells and screening for positive clones with correct insertion was performed by PCR using a forward primer in the genomic Gt(ROSA)26Sor locus 5′ of the targeting vector and a reverse primer in the 5′ region of the targeting vector. Correctly targeted clones showed a 1.3 kb band as a result of the PCR screening. This was further validated by Southern Blot analysis. Three of the positive ES cell clones (1B12, 2D4 and 2B8) were selected for injection into blastocysts from C57BL/6 mice to generate chimeric mice. This part of the procedure was performed by the London Research Institute Transgenic Service. Genotyping of the chimeras was performed using primers to detect the eGFP reporter gene. Germline transmission and line establishment was achieved for clone 1B12. The line is referred to as STOPFloxc-MYC. |
| Phenotypic information | Homozygous:No obvious phenotypes has been observed, overexpression of c-MYC is achieved only after crossing with a cre recombinase-expressing line.Heterozygous:No obvious phenotypes has been observed, overexpression of c-MYC is achieved only after crossing with a cre recombinase-expressing line. |
| Breeding history | STOPFloxc-MYC males have been crossed with C57BL/6 wild-type females to obtain heterozygous offsprings to maintain the line. |
| References |
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| Homozygous fertile | yes |
| Homozygous viable | yes |
| Homozygous matings required | no |
| Immunocompromised | no |
Information from EMMA
| Archiving centre | Mary Lyon Centre at MRC Harwell, Oxford, United Kingdom |
Literature references
- c-MYC overexpression induces choroid plexus papillomas through a T-cell mediated inflammatory mechanism.;Merve Ashirwad, Zhang Xinyu, Pomella Nicola, Acquati Serena, Hoeck Joerg D, Dumas Anaelle, Rosser Gabriel, Li Yichen, Jeyapalan Jennie, Vicenzi Silvia, Fan Qianhai, Yang Zeng Jie, Sabò Arianna, Sheer Denise, Behrens Axel, Marino Silvia, ;2019;Acta neuropathologica communications;7;95; 31142360