B6(129)-H13^{tm1c(KOMP)Wtsi}/HghJ

Stock number: 034743
Product name: SPP^flox
Research areas: Research Tools, Virology Research

Description

SPP^flox mice have loxP sites flanking exon 3 of the histocompatibility 13 (also known as signal peptide peptidase) gene. These mice may be useful in studies of herpes simplex virus infection.

Detailed description

H13 (histocompatibility 13 or signal peptide peptidase - SPP) is member of the γsecretase family and encodes an endoplasmic reticulum protein that cleaves signal peptides after their release by signal peptidase. Complete loss of SPP results in embryonic lethality. SPP is known to bind with herpes simplex virus (HSV) glycoprotein K, and the inducible loss of SPP alters HSV infectivity. SPP^flox mice have loxP sites flanking H13 exon 3. Homozygous mice are healthy, appear normal and are fertile.When bred to mice that express tissue-specific Cre recombinase, resulting offspring will have exon 3 deleted in the Cre-expressing tissues.

For example, when bred to mice with widespread expression of inducible Cre recombinase (Gt(ROSA)26Sor^{tm1(cre/ERT2)Tyj}), tamoxifen-fed progeny infected with HSV-1 in the eye, a model for herpes stromal keratitis, exhibit less HSV-1 replication, a reduction in cellular infiltrates, and a reduced latency of infection.

Development

SPP^flox mice were generated in the laboratory of Dr. Homayon Ghiasi at the Cedars Sinai Medical Center by first obtaining embryonic stem cells  (EPD0867_5_D04) from the International Knockout Mouse Consortium, then generating mutant mice and then breeding to Flpe-deleter mice. Specific details are below.

The L1L2_Bact_P cassette was inserted at position 152680562 of Chromosome 2 upstream of exon 3(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 exon 3 at position 152681394. The construct was electroporated into C57BL/6N-A^tm1Brd derived JM8A3.N1 embryonic stem (ES) cells. Correctly targeted ES cells were injected into blastocysts. The resulting chimeric animals were bred to B6.129S4-Gt(ROSA)26Sor^tm1(FLP1)Dym/RainJ (Stock 009086) females to remove the neomycin cassette. Mice were then bred to B6.129-Gt(ROSA)26Sor^{tm1(cre/ERT2)Tyj}/J (Stock 008463) by the donating lab (see SNP note below), progeny still carrying the loxP-flanked exon 3 allele were selected and the cre allele bred out of the strain. Upon arrival, mice were bred to C57BL/6NJ (Stock No. 005304) for at least one generation to establish the colony.

A 48 SNP (single nucleotide polymorphism) panel analysis, with 43 markers covering all 19 chromosomes and the X chromosome, as well as 5 markers that distinguish between the C57BL/6J and C57BL/6N substrains, was performed on the rederived living colony at The Jackson Laboratory Repository. One of the 43 markers on Chromosome 2 was segregating with 129, likely a passenger segment for H13 mutation. Also, 4 of the 5 markers that determine C57BL/6J from C57BL/6N were found to be segregating. These data suggest the mice sent to The Jackson Laboratory Repository were on a mixed C57BL/6N;C57BL/6J genetic background.

Allele

Symbol: H13^{tm1c(KOMP)Wtsi}
Name: targeted mutation 1c, Wellcome Trust Sanger Institute
MGI accession ID: MGI:6384976
Generation method: Targeted
Attributes: Conditional ready (e.g. floxed)
Synonyms: SPP^flox
Marker symbol: H13
Marker name: histocompatibility 13
Marker synonyms: 1200006O09Rik; 4930443L17Rik; 5031424B04Rik; H-13; Hm13; HM13-IT1; IMP1; IMPAS; IMPAS-1; MSTP086; PSENL3; PSL3; SPP; SPPL1
Chromosome: 2
Strain of origin: C57BL/6N-A^tm1Brd
Molecular note: The L1L2_Bact_P cassette was inserted at position 152522482 of Chromosome 2 upstream of exon 3(Build GRCm39). 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 exon 3 at position 152523314. Flp-mediated recombination removed the FRT-flanked neo cassette leaving exon 3 floxed.