C57BL/6N-Fgd5^tm2Djr/J

Stock number: 027788
Product name: Fgd5-ZsGreen
Research areas: Hematological Research, Research Tools

Description

These Fgd5^ZsGr·ZsGr/+ knock-in/knock-out mice function as a reporter strain and may be suitable for identification and purification of hematopoietic stem cells based on single-color fluorescence.

Detailed description

The Fgd5 gene encodes a protein that upregulates CDC42 activity and is a marker for immunophenotypic hematopoietic stem cells. These mice carry a knock-in/knock-out mutation of the Fgd5 gene in which tandem ZsGreen, Zoanthus sp. green fluorescent protein, cassettes (separated by a 2A peptide), have been inserted in frame into exon 1.

ZsGreen expression is almost exclusively specific to bone marrow hematopoietic stem cells, with some low level of expression detected in bone marrow multipotent progenitor cells. ZsGreen is also expressed in endothelial cells. Mice that are heterozygous for the targeted mutation are viable and fertile. Homozygous mice have an embryonic lethal phenotype, failing to develop past E11.5–E12.0.

Development

A targeting vector was used to insert tandem ZsGreen cassettes separated by a 2A peptide cassette in frame into the first coding exon with a FRT site flanked PGK-NEO cassette downstream of the ZsGreen cassettes. The construct was electroporated into C57BL/6NTac derived B6-3 embryonic stem (ES) cells. Correctly targeted ES cells were injected into blastocysts. The resulting chimeric animals were tested for germline transmission. Upon arrival at The Jackson Laboratory, the mice were crossed to C57BL/6NJ (Stock No. 005304) at least once to establish the colony.

Allele

Symbol: Fgd5^tm2Djr
Name: targeted mutation 2, Derrick Rossi
MGI accession ID: MGI:5642323
Generation method: Targeted
Attributes: Reporter; Null/Knockout
Marker symbol: Fgd5
Marker name: FYVE, RhoGEF and PH domain containing 5
Chromosome: 6
Strain of origin: C57BL/6NTac
Expressed genes: GFP,Green Fluorescent Protein,
Site of expression: ZsGreen expression is detected in all bone marrow blood vessels in adult mice.
Molecular note: The targeting vector consists of tandem ZsGreen cassettes separated by a 2A peptide cassette followed by a FRT-flanked neomycin cassette inserted in frame into exon 1. ZsGreen expression is detected in all bone marrow blood vessels in adult mice by laser-scanning cytometry.