B6;129S4-Zfand5^{Gt(ROSA)72Sor}/J

Stock number: 007203
Research areas: Cell Biology Research, Developmental Biology Research, Endocrine Deficiency Research

Description

These Zfand5-mutant mice may be useful in studying cellular signaling in development and adult mice; specifically receptor tyrosine kinases (RTK; such as Ras, MAP kinase, PI3K and those in the platelet-derived growth factor (PDGF) family) and immediate early genes (IEG) induced shortly after RTK activation. Homozygous embryos exhibit hemorrhages and microaneurisms, with fewer vascular smooth muscle cells found in blood vessels. Skeletal defects are observed in 20% of animals in the sternum and calvarial bones.

Detailed description

Homozygous embryos E11.5 to E18.5 exhibit hemorrhages and microaneurisms. Histological examination of E18.5 homozygous embryos reveals thin blood vessel walls, hemorrhages and lung edema. There are fewer vascular smooth muscle cells (vSMCs) in blood vessels as indicated by immunohistochemistry for desmin and alpha-smooth muscle actin. Skeletal defects are observed in 20% of animals in the sternum and calvarial bones. Homozygotes die a few hours after birth due to difficulty breathing and bruising is visible beneath the skin. Heterozygotes are viable and fertile. No gene product is detected in homozygous embryos aged ED9.5-12.5 or in adult gonad. These Zfand5-mutant mice may be useful in studying cellular signaling in development and adult mice; specifically receptor tyrosine kinases (RTK; such as Ras, MAP kinase, PI3K and those in the platelet-derived growth factor (PDGF) family) and immediate early genes (IEG) induced shortly after RTK activation.

Development

The retroviral gene-trap vector ROSAFARY (or reverse orientation splice acceptor for array) was designed with a promoter trap module (SAβgeo*pA; encoding a lacZ-neo fusion gene) and an frt-flanked poly-A trap module (PGKhygSD; PGK promoter-driven hygromycin gene with adenoviral splice donor). This construct was electroporated into 129S4/SvJaeSor-derived AK7.1 embryonic stem (ES) cells. ES cells with the ROSAFARY vector correctly targeted to the Zfand5 gene (in the 1st intron) were identified and used to generate mutant mice. Heterozygous mice were maintained on a mixed B6;129 background prior to arrival at The Jackson Laboratory. The mice were then crossed to C57BL/6J (Stock No. 000664) for one generation.

The ROSAFARY vector inserts a promoterless lacZ-neo reporter fusion gene (which functions as an artificial 3' terminal exon to intercept and terminate transcription from the targeted gene promoter) as well as a poly-A trap (which functions as an artificial 5' terminal exon to initiate transcription from the insertion site). After inserting into an intron of an endogenous gene at a permissive site and in the correct orientation, the promoter trap module and the poly-A trap module can be activated to form fusion transcripts with the 5' or 3' exons.

Allele

Symbol: Zfand5^{Gt(ROSA)72Sor}
Name: gene trapped 72, Philippe Soriano
MGI accession ID: MGI:3526860
Generation method: Gene trapped
Attributes: Reporter; Null/Knockout
Marker symbol: Zfand5
Marker name: zinc finger, AN1-type domain 5
Chromosome: 19
Strain of origin: 129S4/SvJaeSor
Molecular note: The Rosafary gene trap vector containing a SAbetaGeo pA promoter trap module and a PGKhygSD poly-A trap module was inserted into the first intron of the locus.