B6;Cg-Per1tm1.1Ual/Orl
| Status | Available to order |
| EMMA ID | EM:14846 |
| Citation information | RRID:IMSR_EM:14846 Research Resource Identifiers (RRID) are persistent unique ID numbers assigned to help researchers cite key resources (e.g. antibodies, model organisms and software projects) in the biomedical literature to improve transparency and reproducibility in research. See https://www.rrids.org/ for more information. |
| International strain name | B6;Cg-Per1tm1.1Ual/Orl |
| Alternative name | B6;129P2Per1tm1Ual (Per1 loxP) |
| Strain type | Targeted Mutant Strains : Conditional mutation |
| Allele/Transgene symbol | Per1tm1.1Ual |
| Gene/Transgene symbol | Per1 |
Information from provider
| Provider | Doron Shmerling |
| Provider affiliation | Polygene AG |
| Genetic information | A cassette containing neomycin acetyl-transferase (neo) flanked by two FRT sites was inserted into a bacterial artificial chromosome encompassing the Per1 locus (clone bMQ-339m04). Two loxP sites flanking exon 4-16 of Per1 were introduced to delete exons 4-16 and the reading frame could be resumed in exon 22, thereby deleting almost the entire gene. The neo cassette for selection in ES cells was between exon 3 in the short arm of homology (1.4 kb) and the loxP site upstream of exon 4. The loxP site downstream of exon 16 was followed by the long arm of homology (4 kb) containing exons 17-22. HM-1 ES cells (1x10e7 cells) were electroporated with the linearized targeting vector using standard methods. Gene targeting was performed by PolyGene (Rümlang, Switzerland). Homologous recombination was verified using a primer just outside the short arm of homology of the targeting vector (5’-CTGCCTTTCCTGTCACTATC-3’) and a primer in the neo-cassette (5’-GTTGTGCCCAGTCATAGCCGAATAG-3’) resulting in an amplicon of 2.5 kb. Recombination on the long arm of homology was verified by Southern blotting using the following primers to generate a probe of 490 bp in size: (5’-TGTGGCAGCAGCGTTCAAG-3’) and (5’-GGCGTGGACAATCCTCCAAATG-3’). The probe recognized a 10 kb band for the targeted allele and a 20 kb band for the wild-type allele. C57BL/6 female mice (Charles River, WIGA Sulzfeld) were super-ovulated using standard procedures and mated with C57BL/6 breeder males (Charles River). Blastocysts were injected with ES cell clones and transferred into pseudo-pregnant B6CBAF1 females (Polygene, Rümlang, Switzerland, originally obtained as inbred strains from Harlan Laboratories). Male chimeras were bred with C57BL/6 females. The offspring was then screened for the presence of the neo gene, since the females did not contain the flp recombinase and hence targeted animals still contained the neo cassette. The primers used to detect the neo cassette were: Neo.MP1: 5'-GCTGTGCTCCACGTTGTCAC-3' (neo-cassette and Chromosome 3) Neo.MP5: 5'-GGAAAGCTGGGCTTGCATCTC-3' (Chromosome 3) Neo.MP6: 5'-GGAGCGGCGATACCGTAAAG-3' (neo-cassette) The amplicons for Neo were 512 bp and for wild-type 380 bp. The targeted animals with the neo cassette were then bred with flp deleter mice to remove the neo cassette. Offspring were tested for deletion of the neo cassette using the following primers: F001.1: 5'-GAGCAGGACAACCCATCTAC-3' F001.22: 5'-ACCCTGAACCTGCTTGAC-3' giving rise to a wild-type amplicon of 280 bp and a targeted amplicon of 468 bp. To confirm the presence or the absence of the flp-recombinase we used the following primers: SD24: 5'-CTAATGTTGTGGGAAATTGGAGC-3' SD25: 5'-CTCGAGGATAACTTGTTTATTGC-3' the flp-allele was 568 bp. Screening for the distal loxP site was done with the following primers: F001.23: 5'-GGAGAGCTGCAACATTCC-3' F001.24: 5'-GGAGCTGAAGCTACACTGAC-3' F001.25 (loxP): 5'-ACTAGTTCTAGAGCGGCCGAGC-3' The primer pairs gave rise to the following amplicons: F001.23/F001.24 wild-type allele 476 bp, targeted allele 723 bp. F001.24/F001.25 only detected the targeted allele with an amplicon of 358 bp. For detection of the deleted allele after cre recombinase-mediated recombination we used the primers F001.1 and F001.24 (see above) giving rise to an amplicon of 585 bp. See: 1) Liu P, Jenkins NA, Copeland NG. A highly efficient recombineering-based method for generating conditional knockout mutations. Genome Res. 2003;13(3):476-84. 2) Farley FW, Soriano P, Steffen LS, Dymecki SM. Widespread recombinase expression using FLPeR (flipper) mice. Genesis. 2000;28(3-4):106-10. |
| Phenotypic information | Homozygous:The floxed Per1 animals do not show any phenotype in the homozygous state. Upon deletion of the gene after crossing with a cre recombinase total deleter, the animals display shortened circadian period and increased immobility in the forced swim test. Feeding, breeding and body weight are normal and comparable to controls (Olejniczak et al., PLoS Genet. 2021, 17(7):e1009625).Heterozygous:No phenotype observed. |
| Breeding history | Backcrossed for 2 generations to C57BL/6. |
| References |
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| Homozygous fertile | yes |
| Homozygous viable | yes |
| Homozygous matings required | no |
| Immunocompromised | no |
Information from EMMA
| Archiving centre | CNRS-TAAM – Typing and Archiving of Animal Models, Orléans, France |
| Animals used for archiving | homozygous C57BL/6.129 males |
Literature references
- Light affects behavioral despair involving the clock gene Period 1.;Olejniczak Iwona, Ripperger Jürgen A, Sandrelli Federica, Schnell Anna, Mansencal-Strittmatter Laureen, Wendrich Katrin, Hui Ka Yi, Brenna Andrea, Ben Fredj Naila, Albrecht Urs, ;2021;PLoS genetics;17;e1009625; 34237069