STOCK Hgf^{tm1.1(HGF)Aveo} Prkdc^scid/J

Stock number: 014543
Product name: hHGFki;scid
Research areas: Cancer Research, Cell Biology Research, Hematological Research, Immunology, Inflammation and Autoimmunity Research, Internal/Organ Research, Research Tools, Virology Research

Description

Of note, hHGFki;scid mice also harboring the Il2Rγc^null mutation on the NOD-congenic background are available: NSG-hHGFki (Stock No. 014553).

These hHGFki;scid mice harbor the "humanized" knock-in mutation (hHGFki) that replaces the mouse hepatocyte growth factor (HGF) coding factor, and the Prkdc^scid mutation that results in T- and B-cell deficiency. These mice may be useful in studying the HGF/MET pathway in human tumor xenografts and mouse tumor allografts.

Detailed description

Mice homozygous for both the hHGFki and Prkdc^scid alleles, also called immunocompromised hHGFki mice, are viable and fertile.

The hHGFki allele is a "humanized" knock-in mutation that replaces the mouse hepatocyte growth factor (HGF) coding region downstream of the signal sequence with the human HGF cDNA sequence. As a result, the endogenous mouse promoter drives expression of human HGF. While the human HGF activates both the human and murine form of its tyrosine kinase receptor (met proto-oncogene (MET; c-Met)), the murine HGF is unable to activate human MET. Mice homozygous for hHGFki express only the human form of HGF. In homozygous hHGFki mice, HGF expression from the knock-in allele is observed in developing embryo, as well as adult liver, kidney and lung. hHGFki homozygous mice exhibit an increase in serum HGF after clotting.

Mice homozygous for the Prkdc^scid mutation exhibit T- and B-cell deficiency.

Development

A targeting vector was designed to replace exons 3-6 of the mouse hepatocyte growth factor (Hgf) locus with a cDNA sequence encoding exons 2-18 of the human hepatocyte growth factor (HGF), a polyA signal, and a frt-flanked PGK-neo cassette. The construct was electroporated into 129S6/SvEvTac-derived JB26A10 embryonic stem (ES) cells. Next, ES cells were transiently transfected with a FLPe expressing plasmid to remove the frt-flanked PGK-neo cassette. Correctly targeted ES cells were then injected into recipient blastocysts and chimeric mice were bred with C57BL/6 mice to establish the hHGFki colony. The hHGFki mice were then bred with C3H^scid (C3SnSmn.CB17-Prkdc^scid/J ; see Stock No. 001131) to generate the hHGFki;scid double mutant mice. Mice homozygous for both alleles were bred together for many generations prior to sending to The Jackson Laboratory Repository. Upon arrival, mice were bred together and/or with C3SnSmn.CB17-Prkdc^scid/J ; see Stock No. 001131) for at least one generation to establish the colony. After this, mice homozygous for both alleles were bred together to maintain the colony.

Allele

Symbol: Hgf^{tm1.1(HGF)Aveo}
Name: targeted mutation 1.1, AVEO Pharmaceuticals Inc
MGI accession ID: MGI:4939654
Generation method: Targeted
Attributes: Inserted expressed sequence; Humanized sequence
Marker symbol: Hgf
Marker name: hepatocyte growth factor
Chromosome: 5
Strain of origin: 129S6/SvEvTac
Expressed genes: HGF,hepatocyte growth factor,human
Molecular note: A targeting vector was designed to replace exons 3-6 of the mouse hepatocyte growth factor (Hgf) locus with a cDNA sequence encoding exons 2-18 of the human hepatocyte growth factor (HGF), a polyA signal, and a frt-flanked PGK-neo cassette. Flp-mediated recombination removed the neo cassette.
General note: ES cells = JB26A10 (129S6/SvEvTac)

Allele

Symbol: Prkdc^scid
Name: severe combined immunodeficiency
MGI accession ID: MGI:1857113
Generation method: Spontaneous
Marker symbol: Prkdc
Marker name: protein kinase, DNA activated, catalytic polypeptide
Chromosome: 16
Strain of origin: C.BKa-Igh^b/Icr
Site of expression: T and B lymphocytes.
Molecular note: A T-to-A transversion point mutation at a position corresponding to codon 4046 (codon 4095 in transcript ENSMUST00000023352.8) created a premature stop codon (p.Y4046*).