B6;129P2-Hprttm1(CAG-mCherry-Top1)Jtu/H
| Status | Available to order |
| EMMA ID | EM:15010 |
| International strain name | B6;129P2-Hprttm1(CAG-mCherry-Top1)Jtu/H |
| Alternative name | Hprttm1(CAG-mCherry Top1)Jtu |
| Strain type | Targeted Mutant Strains : Knock-in |
| Allele/Transgene symbol | Hprttm1(CAG-mCherry-Top1)Jtu |
| Gene/Transgene symbol | Hprt1 |
Information from provider
| Provider | James Turner |
| Provider affiliation | Sex Chromosome Biology Laboratory, The Francis Crick Institute |
| Additional owner | Genoway, FRANCE |
| Genetic information | The mice harbour an sgRNA transgene targeting Top1 gene which has been inserted into Hprt1 locus. The Top1 sgRNA targeting vector was generated using plasmid pX333 as a backbone (Addgene). The Cas9 cassette was replaced with an mCherry reporter. Top1 sgRNA2 were inserted using BbsI. The resulting hU6-sgRNA2-pCbh-mCherry construct was used to generate the XTop1 transgene by GenOway (Lyon, France) using their Quick Knock-in approach in E14Tg2a mESCs derived from 129P2/OlaHsd. Human HPRT1 exons 1 and 2 were inserted into the sgRNA2-mCherry targeting vector. The construct was targeted to the Hprt1 locus in mESCs that lacked exons 1 and 2. Upon recombination of the targeting vector into Hprt1, gene function was restored. Correctly targeted clones were selected with hypoxanthine-aminopterin-thymidine (HAT) medium. Eight HAT-resistant clones were selected and amplified for confirmation of on-target integration. X-integration clones were confirmed using PCR and Sanger sequencing for validation. |
| Phenotypic information | Homozygous:Unknown. Homozygous mice were not generated for our experiments.Heterozygous:Viable and fertile. |
| Breeding history | 4 generations of backcrosses to C57BL/6 |
| References |
|
| Homozygous fertile | not known |
| Homozygous viable | not known |
| Homozygous matings required | no |
| Immunocompromised | no |
Information from EMMA
| Archiving centre | Mary Lyon Centre at MRC Harwell, Oxford, United Kingdom |
Literature references
- CRISPR-Cas9 effectors facilitate generation of single-sex litters and sex-specific phenotypes.;Douglas Charlotte, Maciulyte Valdone, Zohren Jasmin, Snell Daniel M, Mahadevaiah Shantha K, Ojarikre Obah A, Ellis Peter J I, Turner James M A, ;2021;Nature communications;12;6926; 34862376