B6J.129P2-Pofut2^{Gt(RST434)Byg}/BchMmjax

Stock number: 037014
Research areas: Developmental Biology Research, Research Tools

Description

This RST434 gene trap mutation abolishes endogenous Pofut2 (protein O-fucosyltransferase 2) gene function while expressing a β-geo reporter fusion protein from the disrupted locus. Homozygous deletion of Pofut2 results in embryonic lethality. This strain may be useful for studying the role of O-fucose in gastrulation.

Detailed description

POFUT2 (protein O-fucosyltransferase 2) functions to add O-fucose to serine or threonine residues in thrombospondin type 1 repeats. The RST434 gene trap mutation abolishes O-fucosyltransferase activity while expressing a β-geo reporter fusion protein from the disrupted locus. Homozygous deletion of Pofut2 results in embryonic lethality during gastrulation. At the onset of gastrulation E6.5 embryos appear to have a rounder appearance, a shortened proximal-distal axis, a thickened and disorganized endoderm (embryonic and extra-embryonic) and constriction in the junction between the extra-embryonic ectoderm and ectoplacental cone. No homozygous embryos are found beyond E7.5. Heterozygotes are viable and fertile. Mutant mice may be useful as a lacZ reporter for Pofut2 expression or as a knockout model for studying the role of O-fucose in gastrulation.

Development

Bay Genomics gene trap stem cell clone RST434 contains a splice acceptor site, β-geo (a fusion of β-galactosidase and neomycin phosphotransferase II), an IRES, an hPLAP and a polyadenylation site inserted into exon 6 of the Pofut2 gene on chromosome 10. The insertion results in a fusion between exons 1-5 and the β-geo and creates a functionally null allele. The construct was electroporated into 129P2/OlaHsd derived E14TG2a.4 embryonic stem (ES) cells. Correctly targeted ES cells were injected into recipient blastocysts. The resulting chimeric animals were crossed to C57BL/6J mice and backcrossed for 31 generations by the donating laboratory. Upon arrival, sperm was cryopreserved. To establish a live colony, an aliquot of frozen sperm is used to fertilize C57BL/6J.

Allele

Symbol: Pofut2^{Gt(RST434)Byg}
Name: gene trap RST434, BayGenomics
MGI accession ID: MGI:4127173
Generation method: Gene trapped
Attributes: Reporter; Null/Knockout
Synonyms: RST434
Marker symbol: Pofut2
Marker name: protein O-fucosyltransferase 2
Marker synonyms: 2310011G23Rik; C21orf80; FUT13; MGC:6943
Chromosome: 10
Strain of origin: 129P2/OlaHsd
Molecular note: A gene trap vector insertion occurred within exon 6 and resulted in an inframe fusion between Pofut2 (exons 1 through 5) and beta-gal-neomycin. The resulting fusion protein lack TSR-specific O-fucosyltransferase activity. It is a null allele.