B6N(129S4)-Srd5a3^{tm1c(EUCOMM)Wtsi}/J

Stock number: 034731
Product name: Srd5a3^fl
Research areas: Cell Biology Research, Research Tools

Description

Srd5a3^fl mice have loxP sites flanking exons 2-4 of the steroid 5 alpha-reductase 3 gene. Srd5a3 encodes a polyprenol reductase required in N-linked glycosylation.

Detailed description

Srd5a3^fl mice have loxP sites flanking exons 2-4 of the steroid 5 alpha-reductase 3 gene. Srd5a3 encodes a polyprenol reductase required in N-linked glycosylation. As phenotype data is generated it will be made available to the public.

Development

The L1L2_Bact_P cassette was inserted at a position on Chromosome 5 upstream of exon 2 of the steroid 5 alpha-reductase 3 ( Srd5a3) gene. The cassette is composed of a frt site followed by lacZ sequence and a loxP site. This first loxP site is followed by neomycin under the control of the human beta-actin promoter, SV40 polyA, a second frt site and a second loxP site. A third loxP site is inserted downstream of exon 4. The critical exons, 2-4, are thus flanked by loxP sites. The construct was introduced into C57BL/6N-derived JM8.N4 embryonic stem (ES) cells, and correctly targeted ES cells were injected into B6(Cg)-Tyr^c-2J/J (Stock No. 000058) blastocysts. The resulting chimeric males were bred to C57BL/6NJ (Stock No. 005304) females. Frozen sperm was imported to The Jackson Laboratory. To establish our live colony, an aliquot of frozen sperm was used to fertilize oocytes from B6N.ROSA26::FLPe mice (Stock No. 016226) to remove the frt-flanked neomycin and lacZ sequences from resulting offspring. Progeny were subsequently bred to C57BL/6NJ mice to remove the FLPe-expressing transgene.

Allele

Symbol: Srd5a3^{tm1c(EUCOMM)Wtsi}
Name: targeted mutation 1c, Wellcome Trust Sanger Institute
MGI accession ID: MGI:6295430
Generation method: Targeted
Attributes: Conditional ready (e.g. floxed)
Marker symbol: Srd5a3
Marker name: steroid 5 alpha-reductase 3
Chromosome: 5
Strain of origin: C57BL/6N-A^tm1Brd
Molecular note: The L1L2_Bact_P cassette was inserted at position 76147218 of Chromosome 5 upstream of the critical exon(s) (Build GRCm38). The cassette is composed of an FRT site followed by lacZ sequence and a loxP site. This first loxP site is followed by a neomycin resistance gene under the control of the human beta-actin promoter, SV40 polyA, a second FRT site and a second loxP site. A third loxP site is inserted downstream of the targeted exon(s) at position 76150215. The critical exon(s) is/are thus flanked by loxP sites. A "conditional ready" (floxed) allele was created by flp recombinase expression in mice carrying this allele to remove the lacZ sequence and neo selection cassette, leaving loxP sites flanking the critical exon(s). Further information on targeting strategies used for this and other IKMC alleles can be found at http://www.informatics.jax.org/mgihome/nomen/IKMC_schematics.shtml