B6.129S6-Pask^tm1Weng/J

Stock number: 017709
Research areas: Diabetes and Obesity Research, Metabolism Research, Reproductive Biology Research, Research Tools

Description

The Pask gene is disrupted by the insertion of an IRES-β-geo fusion cassette in these knockin/knockout mice. LacZ is detected in testis and homozygotes exhibit a range of metabolic traits.

Detailed description

A splice acceptor site and an IRES-β-geo fusion cassette replace exons 10-14 of the PAS domain containing serine/threonine kinase (Pask) gene in these mutant mice. Gene function is abolished. PASK is a serine/threonine kinase which regulates energy homeostasis and acts as a cellular metabolic sensor. In wildtype mice, PASK is expressed in post-meiotic germ cells during spermatogenesis, with typically low level ubiquitous expression. Homozygous Pask^-/- mice are viable, fertile, and normal in size, with lacZ staining visible in the testis beginning at 25 days of age. These mice exhibit impaired glucose-stimulated insulin secretion with no change in islet size or morphology. Increased O₂ consumption, CO₂ production, and heat generation is also observed. On a high-fat diet, Pask^-/- mice have reduced liver triglyceride levels and show little difference in glucose tolerance, insulin resistance, or weight gain compared to wildtype mice.

Development

A targeting vector was designed to replace exons 10-14 of the PAS domain containing serine/threonine kinase (Pask) gene with a splice acceptor site, an internal ribosome entry site (IRES), and a neomycin-β-galactosidase fusion protein (β-geo). The construct was electroporated into 129S6/SvEvTac-derived TC-1 embryonic stem (ES) cells. Correctly targeted ES cells were injected into (C57BL/6 x DBA)F1 blastocysts and the resulting chimeric males were bred to C57BL/6 females. These mice were backcrossed at least 10 generations to C57BL/6J background. Upon arrival at The Jackson Laboratory, mice were bred to C57BL/6J (Stock No. 000664) for at least one generation to establish the colony.

Allele

Symbol: Pask^tm1Weng
Name: targeted mutation 1, Roland H Wenger
MGI accession ID: MGI:2677908
Generation method: Targeted
Attributes: Reporter; Null/Knockout
Marker symbol: Pask
Marker name: PAS domain containing serine/threonine kinase
Chromosome: 1
Strain of origin: 129S6/SvEvTac
Expressed genes: Bgeo,fusion of beta-galactosidase and neomycin phosphotransferase genes,E. coli
Site of expression: lacZ staining visible in the testis beginning at 25 days of age.
Molecular note: Exons 10 through 14 were replace with a cassette containing a splice acceptor site and IRES-beta-geo. Norhtern blot analysis of various tissues confirmed the absence of endogenous transcript and identified mutant transcript containing the beta-geo squenence.