B6.129(Cg)-Msi2^tm1.1Geno/ReyaJ

Stock number: 035371
Product name: Msi2^eGFP
Research areas: Cancer Research, Cell Biology Research, Research Tools

Description

Msi2^eGFP knock-in/knock-out mice have an enhanced green fluorescent protein (EGFP) sequence inserted into exon 1 of the Msi2 gene. These mice can be used to identify and track normal stem cells, cancer stem cells, and chemotherapy and radiotherapy-resistant cancer cells in vivo, and may help with studies of normal and cancer stem cell function as well as cancer diagnostic and therapeutic studies.

Detailed description

Msi2^eGFP knock-in/knock-out mice have an enhanced green fluorescent protein (EGFP) sequence and a STOP codon inserted into exon 1 of the Musashi RNA-binding protein 1 (Msi2) gene. Msi2 encodes an RNA binding protein that plays an important role in post-transcriptional gene regulation. It is associated with stem and progenitor cells in different tissues, and found to be essential for cell fate determination in context of the nervous and hematopoietic systems. Msi2 is highly upregulated in many aggressive cancers, including myeloid leukemia and pancreatic cancer, and is a poor prognostic factor for outcome. Insertion of the reporter gene abolishes expression of Msi2. Mice that are heterozygous for this knock-in/knock-out allele are viable and fertile. EGFP can be detected by direct fluorescence or through an anti-GFP antibody. The Msi2^eGFP reporter mice can be used to identify and track normal stem cells, cancer stem cells, and chemotherapy and radiotherapy-resistant cancer cells in vivo, and may help with studies of normal and cancer stem cell function as well as cancer diagnostic and therapeutic studies.

Heterozygous mice are viable and fertile, the donating lab has not attempted to make a homozyogus colony.

Development

The Msi2^eGFP targeting vector was designed to insert an enhanced green fluorescent protein (EGFP) sequence, followed by a STOP cassette and a loxP-flanked neomycin resistance (neo) cassette, into exon 1 of the musashi RNA-binding protein 2 (Msi2) gene. The construct was electroporated into 129Sv-derived embryonic stem (ES) cells and correctly targeted ES cells were injected into recipient blastocysts. The resulting chimeric mice were bred with cre recombinase expressing mice to remove the neo cassette, and resulting mice were subsequently bred together to remove the cre-expressing transgene. Offspring were further bred to C57BL/6J mice for at least 10 generations. Upon arrival at The Jackson Laboratory, sperm was cryopreserved. To establish our live colony, an aliquot of frozen sperm was used to fertilize C57BL/6J oocytes (Stock No. 000664).

Allele

Symbol: Msi2^tm1.1Geno
Name: targeted mutation 1.1, Genoway
MGI accession ID: MGI:6324502
Generation method: Targeted
Attributes: Reporter; Null/Knockout
Synonyms: Msi2^GFP
Marker symbol: Msi2
Marker name: musashi RNA-binding protein 2
Marker synonyms: MSI2H; Musashi2; RGD1560397
Chromosome: 11
Strain of origin: 129
Site of expression: Normal and cancer stem cells.
Molecular note: The targeting vector is designed to insert an enhanced green fluorescent protein (EGFP) sequence, followed by a STOP cassette and a loxP-flanked neomycin resistance (neo) cassette, into exon 1.Cre-mediated recombination removed the floxed neo cassette.