IMPC phenotypes (gene matching)
C57BL/6-Tyrc-Brd Hprt1em2(CAG-cas9,-EGFP)Jtu/H
| Status | Available to order |
| EMMA ID | EM:15012 |
| Citation information | RRID:IMSR_EM:15012 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 | C57BL/6-Tyrc-Brd Hprt1em2(CAG-cas9,-EGFP)Jtu/H |
| Alternative name | Hprt-em2(Cas9 eGFP)Jtu |
| Strain type | Spontaneous |
| Allele/Transgene symbol | Tyrc-Brd, Hprt1em2(CAG-cas9,-EGFP)Jtu |
| Gene/Transgene symbol | Tyr, Hprt1 |
Information from provider
| Provider | James Turner |
| Provider affiliation | Sex Chromosome Biology Laboratory, The Francis Crick Institute |
| Genetic information | XCas9 mice express Cas-9 linked to EGFP under the control of CAG promoter from the Hprt1 site of the mouse X chromosome. The expression of Cas9 and eGFP is linked via a T2A peptide under a CMV early enhancer/chicken beta-actin (CAG) promoter. The Cas9-eGFP targeting vector was generated using the pX330 plasmid backbone, containing a pCAG driven 3X FLAG-NLS-Cas9-T2A-eGFP construct. X chromosome homology arms, amplified from C57BL/6J DNA, and a LoxP-flanked pPGK-neomycin cassette were inserted using directional cloning or Gibson Assembly. C57BL/6N mouse ES cells (mESCs) were maintained in serum/LIF conditions and transfected with the Cas9-eGFP targeting vector plasmid and an sgRNA targeting Hprt1 exon 2 using Lipofectamine 2000, according to manufacturer’s instructions. Targeted mESC clones were selected by G418 (270 mg/ml) for 8–10 days. Forty-eight surviving clones were picked into a 96-well plate and expanded. PCR genotyping was performed on extracted DNA in a total volume of 25 μl (12.5 μl NEB Q5 High-Fidelity Master Mix, 10 mM each primer), using primer forward and reverse pairs aligning to the endogenous Hprt1 locus and to the transgene construct. Resultant PCR amplicons were analysed by gel electrophoresis for corresponding to the expected amplicon size, and by Sanger sequencing. Of 48 clones, 9 were found to be successfully targeted (19%). Targeted mESC clones were injected into albino C57BL/6J-Tyrc-Brd blastocysts and surgically transferred into pseudopregnant females. XCas9Y mESC contribution to founders was assessed by coat colour. High contribution transgenic males were bred with C57BL/6J-Tyrc-Brd albino females, and offspring with black coat colour were genotyped for the transgene to confirm germline transmission. |
| Phenotypic information | Homozygous:Viable and fertile with no health concerns.Heterozygous:Viable and fertile with no health concerns. |
| Breeding history | Mated to C57BL/6J for one generation, then brother x sister matings for 5 generations, then finally mated 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 | Mary Lyon Centre at MRC Harwell, Oxford, United Kingdom |
Disease and phenotype information
Orphanet associated rare diseases, based on orthologous gene matching
- Temperature-sensitive oculocutaneous albinism type 1 / Orphanet_352737
- Oculocutaneous albinism type 1A / Orphanet_79431
- Oculocutaneous albinism type 1B / Orphanet_79434
- Minimal pigment oculocutaneous albinism type 1 / Orphanet_352734
- Ocular albinism with congenital sensorineural deafness / Orphanet_352740
MGI phenotypes (gene matching)
- decreased hematocrit / MGI
- increased leukocyte cell number / MGI
- increased neutrophil cell number / MGI
- abnormal small intestine morphology / MGI
- abnormal liver morphology / MGI
- abnormal branching of the mammary ductal tree / MGI
- enlarged spleen / MGI
- spleen hyperplasia / MGI
- enlarged lymph nodes / MGI
- tremors / MGI
- convulsive seizures / MGI
- abnormal lung morphology / MGI
- decreased body weight / MGI
- decreased anxiety-related response / MGI
- ataxia / MGI
- hypoactivity / MGI
- impaired coordination / MGI
- abnormal gait / MGI
- short stride length / MGI
- decreased exploration in new environment / MGI
- limb grasping / MGI
- abnormal motor coordination/balance / MGI
- abnormal hematopoietic system physiology / MGI
- hyperglycemia / MGI
- anemia / MGI
- cardiac hypertrophy / MGI
- increased mammary adenocarcinoma incidence / MGI
- abnormal reflex / MGI
- seizures / MGI
- abnormal motor capabilities/coordination/movement / MGI
- premature death / MGI
- abnormal definitive hematopoiesis / MGI
- abnormal brain morphology / MGI
- no abnormal phenotype detected / MGI
- neurodegeneration / MGI
- abnormal spleen white pulp morphology / MGI
- abnormal hematopoietic system morphology/development / MGI
- abnormal megakaryocyte progenitor cell morphology / MGI
- hepatic steatosis / MGI
- decreased vertical activity / MGI
- increased heart weight / MGI
- increased systemic arterial blood pressure / MGI
- albuminuria / MGI
- decreased erythrocyte cell number / MGI
- increased urine protein level / MGI
- impaired social transmission of food preference / MGI
- no phenotypic analysis / MGI
- phenotypic reversion / MGI
- abnormal dopaminergic neuron morphology / MGI
- astrocytosis / MGI
- abnormal depression-related behavior / MGI
- decreased tumor growth/size / MGI
- abnormal nervous system morphology / MGI
- abnormal cardiac muscle relaxation / MGI
- neuronal intranuclear inclusions / MGI
- abnormal myocardial fiber physiology / MGI
- abnormal Paneth cell morphology / MGI
- decreased B cell number / MGI
- decreased cardiac muscle contractility / MGI
- glomerulosclerosis / MGI
- abnormal podocyte morphology / MGI
- muscle phenotype / MGI
- homeostasis/metabolism phenotype / MGI
- endocrine/exocrine gland phenotype / MGI
- behavior/neurological phenotype / MGI
- immune system phenotype / MGI
- taste/olfaction phenotype / MGI
- hematopoietic system phenotype / MGI
- jerky movement / MGI
- thrombocytosis / MGI
- decreased ventricle muscle contractility / MGI
- decreased mean corpuscular hemoglobin concentration / MGI
- decreased dopamine level / MGI
- abnormal podocyte slit diaphragm morphology / MGI
- absent podocyte slit diaphragm / MGI
- podocyte foot process effacement / MGI
- increased megakaryocyte cell number / MGI
- abnormal spatial reference memory / MGI
- abnormal spatial working memory / MGI
- abnormal splenic cell ratio / MGI
- abnormal physiological response to xenobiotic / MGI
- abnormal enterocyte proliferation / MGI
- abnormal enterocyte apoptosis / MGI
- abnormal neuron differentiation / MGI
- increased mammary gland tumor incidence / MGI
- myeloid hyperplasia / MGI
- expanded mesangial matrix / MGI
- mesangial cell hyperplasia / MGI
- abnormal habituation to a new environment / MGI
- abnormal ceramide level / MGI
- decreased brain choline acetyltransferase activity / MGI
- decreased brain tyrosine 3-monooxygenase activity / MGI
- decreased vascular endothelial cell proliferation / MGI
- abnormal cell morphology / MGI
- diluted coat color / MGI
- irregular coat pigmentation / MGI
- belly spot / MGI
- absent hair follicle melanin granules / MGI
- shiny fur / MGI
- mottled coat / MGI
- abnormal retinal photoreceptor morphology / MGI
- pigmentation phenotype / MGI
- absent skin pigmentation / MGI
- abnormal keratinocyte apoptosis / MGI
- abnormal eye pigmentation / MGI
- abnormal retina morphology / MGI
- retinal degeneration / MGI
- decreased retinal photoreceptor cell number / MGI
- abnormal coat appearance / MGI
- male infertility / MGI
- abnormal coat/hair pigmentation / MGI
- prenatal lethality / MGI
- premature death / MGI
- abnormal vision / MGI
- abnormal skin pigmentation / MGI
- no abnormal phenotype detected / MGI
- no phenotypic analysis / MGI
- abnormal cell nucleus morphology / MGI
- failure of zygotic cell division / MGI
- single kidney / MGI
- absent seminal vesicle / MGI
- abnormal chromosome morphology / MGI
- chromosome breakage / MGI
- induced chromosome breakage / MGI
- increased cellular sensitivity to ionizing radiation / MGI
- abnormal miscarriage rate / MGI
- abnormal hair follicle melanogenesis / MGI
- abnormal melanosome morphology / MGI
- abnormal iris pigmentation / MGI
- absent coat pigmentation / MGI
- decreased eye pigmentation / MGI
- abnormal aqueous drainage system morphology / MGI
- abnormal retinal ganglion layer morphology / MGI
- abnormal eye physiology / MGI
- abnormal intraocular pressure / MGI
- variegated coat color / MGI
- homeostasis/metabolism phenotype / MGI
- reproductive system phenotype / MGI
- vision/eye phenotype / MGI
- hypopigmentation / MGI
- ocular albinism / MGI
- absent eye pigmentation / MGI
- decreased survivor rate / MGI
- transverse fur striping / MGI
- mortality/aging / MGI
- abnormal survival / MGI
- integument phenotype / MGI
- neonatal lethality, complete penetrance / MGI
- perinatal lethality, complete penetrance / MGI
- prenatal lethality, complete penetrance / MGI
- embryonic lethality, complete penetrance / MGI
- embryonic lethality at implantation, complete penetrance / MGI
- embryonic lethality before implantation, complete penetrance / MGI
- decreased ear pigmentation / MGI
- variegated eye pigmentation pattern / MGI
- decreased a wave amplitude / MGI
- decreased b wave amplitude / MGI
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