B6N.129S6-Sphk2^tm1Rlp/J

Stock number: 019140
Product name: SphK2^-
Strain synonyms: SK2-
Research areas: Apoptosis Research, Cell Biology Research, Immunology, Inflammation and Autoimmunity Research

Description

These knockout/knockin mice express β-galactosidase (lacZ) from a disrupted Sphk2 gene and exhibit a mild anaphylaxis when challenged. They maybe useful for studying the role of sphingosine-1-phosphate in cell survival and proliferation.

Detailed description

An internal ribosome entry site (IRES) followed by a β-galactosidase (lacZ)-neomycin resistance (neo) cassette replaces part of exon 4 and exons 5-7 of the sphingosine kinase 2 (Sphk2) gene, abolishing gene function. SPHK2, expressed in many cell types, is an enzyme that catalyzes the synthesis of sphingosine-1-phosphate (S1P), a lipid signaling molecule, from sphingosine and ATP. S1P regulates immune cell trafficking and vascular function. Mice homozygous for this allele are viable and fertile. SPHK2 KO mice exhibit mild anaphylaxis when challenged, but recover quickly. When bred to SPHK1 KO mice (Stock No. 019095) resulting double knockout mice die by E13.5 due to developmental defects and hemorrhaging.

Development

A targeting vector was designed to replace part of exon 4 and exons 5-7 of the sphingosine kinase 2 (Sphk2) gene with an internal ribosome entry site (IRES) followed by a β-galactosidase (lacZ)-neomycin resistance (neo) cassette. The construct was electroporated into 129S6/SvEvTac-derived TC1 embryonic stem (ES) cells. Correctly targeted ES cells were injected into C57BL/6 blastocysts and the resulting chimeric males were bred to C57BL/6NCrl females. These mice were backcrossed to C57BL/6NCrl mice for at least 7 generations. Upon arrival at The Jackson Laboratory, mice were bred to C57BL/6NJ (Stock No. 005304) for at least one generation to establish the colony.

Allele

Symbol: Sphk2^tm1Rlp
Name: targeted mutation 1, Richard L Proia
MGI accession ID: MGI:3611080
Generation method: Targeted
Attributes: Reporter; Null/Knockout
Marker symbol: Sphk2
Marker name: sphingosine kinase 2
Chromosome: 7
Strain of origin: 129S6/SvEvTac
Expressed genes: lacZ,beta-galactosidase,E. coli
Molecular note: A lacZ-neo cassette replaced half of exon 4 through the end of exon 7 by homologous recombination. RT-PCR analysis of total RNA form adult tissues failed to detect transcript in mutants.