STOCK Pik3c3^{tm1c(EUCOMM)Wtsi}/J

Stock number: 019081
Product name: Vsp34^f
Research areas: Cell Biology Research, Research Tools

Description

Vps34^f/f floxed mice are used to conditionally delete Pik3c3, making them useful for studies of autophagy and exocytosis.

Detailed description

These Vps34^f/f mutant mice possess loxP sites flanking exon 4 of the phosphoinositide-3-kinase, class 3 (Pik3c3) gene. Also known as Vsp34, Pik3c3 generates phosphatidylinositol 3-phosphate, a regulator of membrane trafficking such as autophagy and endocytosis. Mice that are homozygous for this allele are viable and fertile. When these mutant mice are bred to mice that express Cre recombinase, resulting offspring will have exon 4 deleted in the cre-expressing tissues.

For example, when bred to B6.Cg-Tg(Alb-cre)21Mgn/J mice (Stock No. 003574) expressing cre recombinase in liver, resulting mice develop hepatomegaly and hepatic steatosis, with impaired protein turnover. These double mutant mice exhibit decreased survival, with 50% lethality in the first 2 months of life.

For example, when bred to B6.FVB(129S4)-Tg(Ckmm-cre)5Khn/J mice (Stock No. 006475) expressing cre recombinase in heart, mice exhibit cardiomegaly and decreased contractility. These double mutant mice die between 5 and 13 weeks of age.

Development

A targeting vector was designed to insert, from 5' to 3', a frt-site, a β-galactosidase cassette, a neomycin resistance cassette, a second frt-site, and a loxP site upstream of exon 4 of the phosphoinositide-3-kinase, class 3 (Pik3c3) gene. A second loxP site was also inserted just downstream of exon 4 of the targeted gene. The construct was electroporated into C57BL/6N-derived JM8.F6 embryonic stem (ES) cells, and correctly targeted ES cells were injected into blastocysts. Resulting chimeric males were bred to C57BL/6 females, and their offspring were bred to FLP-expressing mice to delete the β-gal and neo cassette. These conditional Vps34^f/f mice were maintained on a mixed background. 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: Pik3c3^{tm1c(EUCOMM)Wtsi}
Name: targeted mutation 1c, Wellcome Trust Sanger Institute
MGI accession ID: MGI:5316387
Generation method: Targeted
Attributes: Conditional ready (e.g. floxed)
Synonyms: Vps34^f
Marker symbol: Pik3c3
Marker name: phosphatidylinositol 3-kinase catalytic subunit type 3
Marker synonyms: 5330434F23Rik; hVps34; Vps34
Chromosome: 18
Strain of origin: C57BL/6N
Molecular note: An FRT-flanked promoterless cassette, which contains a splice acceptor, beta-geo and polyadenylation sequence, with a 3' loxP site was inserted upstream of exon 4. An additional loxP site was inserted downstream of exon 4. Flp-mediated recombination removed the promoterless cassette and left exon 4 floxed.
General note: Cell line EPD0103_4_A07 was used to make chimeric mice. Germline transmission was accomplished and subsequenct recombination established the allele. J:182628