STOCK Kitl^tm1.1Sjm/J

Stock number: 017860
Product name: Scf (Kitl) knockin
Research areas: Cardiovascular Research, Developmental Biology Research, Hematological Research, Internal/Organ Research, Research Tools

Description

An EGFP reporter disrupts expression of the Kitl gene in this targeted mutation strain. EGFP is primarily expressed by perivascular endothelial and stromal cells throughout the bone marrow in heterozygotes.

Detailed description

An Enhanced Green Fluorescent Protein (EGFP) reporter disrupts expression of Kitl (kit ligand; also called stem cell factor, Scf) in this targeted mutation strain. EGFP is primarily expressed by perivascular endothelial and stromal cells throughout the bone marrow. Homozygotes die perinatally with severe anemia. Kitl transcripts are nearly undetectable by quantitatve RT-PCR in the livers of newborn homozygotes. Overall cellularity of the newborn liver is reduced fivefold in homozygotes and approximately twofold in heterozygotes. Hematopoietic stem cell numbers are reduced about eightfold in the livers of newborn homozygotes as compared with wildtype and heterozygous controls. Heterozygotes have a diluted coat color and often have a white spot on the abdomen.

Development

An EGFP-polyA-Frt-Neo-Frt cassette was introduced to the second exon of the targeted gene via 129S6/SvEvTac-derived W4 embryonic stem (ES) cells. Chimeric mice were bred with C57BL/Ka mice to obtain germline transmission, then bred with C57BL/Ka background mice expressing Flp to excise the neomycin cassette. This strain was backcrossed for 10 generations to C57BL/Ka by the donating laboratory. Upon arrival at The Jackson Laboratory, C57BL/6J was used for rederivation.

Allele

Symbol: Kitl^tm1.1Sjm
Name: targeted mutation 1.1, Sean J Morrison
MGI accession ID: MGI:5300819
Generation method: Targeted
Attributes: Reporter; Null/Knockout
Marker symbol: Kitl
Marker name: kit ligand
Chromosome: 10
Strain of origin: 129S6/SvEvTac
Expressed genes: GFP,Green Fluorescent Protein,
Site of expression: EGFP is primarily expressed by perivascular endothelial and stromal cells throughout the bone marrow.
Molecular note: A targeting vector was designed to replace exon 2 with EGFP (in frame) and an FRT-flanked neomycin resistance cassette. The neomycin resistance cassette was removed by flp-mediated recombination. qRT-PCR confirmed the absence of transcript expression in the liver.