STOCK Gt(ROSA)26Sor^{tm16(CAG-Nsd2,-EBFP)Rsky}/J

Stock number: 039717
Product name: MMSET
Strain synonyms: R26 MMSET^stopF
Research areas: Cancer Research, Research Tools

Description

These MMSET mice express Nsd2 and eBFP after removal of the loxP-flanked STOP cassette via Cre-mediated recombination. These mice are a useful tool for developing preclinical models of multiple myeloma (MM).

Of note, the donating laboratory has also made available their Ccnd1 mice (Stock No. 039718), a similar strain with Cre-inducible Ccnd1 and eBFP expression.

Detailed description

These MMSET mice express (from 5' to 3') a loxP-flanked STOP cassette containing a neomycin resistance (neo) cassette, a nuclear receptor binding SET domain protein 2 (Nsd2) cDNA, and a frt-flanked cassette containing an internal ribosome entry site (IRES) and an enhanced blue fluorescent protein (eBFP) sequence, all under control of a CAG promoter from the endogenous Gt(ROSA)26Sor locus. NSD2 encodes the histone methyltransferase, MMSET, which is activated in multiple myeloma (MM), a fatal malignancy of bone marrow plasma cells which is initiated by genetic aberrations in germinal center (GC) B cells.

Homozygous MMSET mice are viable and fertile. In the absence of Cre, Nsd2 and eBFP expression from the Gt(ROSA)26Sor locus is prevented by the STOP cassette. After removal of the loxP-flanked STOP cassette via Cre mediated recombination, Nsd2 and eBFP are expressed. When bred to Cγ1-cre mice (Stock No. 010611), expressing Cre in germinal center B cells, the expression of MMSET can lead to an asymptomatic pre-malignant monoclonal gammopathy of undetermined significance (MGUS) state which involves outgrowth of clonal restricted plasma cells with age.

EBFP fluorescence is detectable by immunofluorescence and FACS analysis. No EBFP fluorescence is detected prior to Cre recombination.

Of note, Ccnd1 mice (Stock No. 039718) are a similar strain with conditional Ccnd1 and eBFP expression.

Development

A targeting vector was designed with (from 5' to 3') a ubiquitously expressed CAG promoter, a loxP-flanked STOP cassette containing a neomycin resistance (neo) cassette, a nuclear receptor binding SET domain protein 2 (Nsd2) cDNA, and a frt-flanked cassette containing an internal ribosome entry site (IRES) and an enhanced blue fluorescent protein (eBFP) sequence. This targeting vector was inserted into the Gt(ROSA)26Sor locus via electroporation into C57BL/6NTac-derived B6-3 embryonic stem (ES) cells. Correctly targeted ES cells were injected into blastocysts and resulting chimeric mice were subsequently bred with other transgenic mice with different genetic backgrounds. The donating laboratory reports the resulting MMSET mice were further bred to C57BL/6N mice for at least 3 generations (to remove the contaminating alleles) prior to sending males to The Jackson Laboratory in 2024. Upon arrival, sperm was cryopreserved. To establish our live colony, an aliquot of frozen sperm was used to fertilize C57BL/6NJ oocytes (Stock No. 005304).

Allele

Symbol: Gt(ROSA)26Sor^{tm16(CAG-Nsd2,-EBFP)Rsky}
Name: targeted mutation 16, Klaus Rajewsky
MGI accession ID: MGI:7703662
Generation method: Targeted
Attributes: Conditional ready (e.g. floxed); Reporter; Inserted expressed sequence
Synonyms: R26 MMSET^stopF
Marker symbol: Gt(ROSA)26Sor
Marker name: gene trap ROSA 26, Philippe Soriano
Marker synonyms: beta geo; Gtrgeo26; Gtrosa26; R26; ROSA26; SETD5-AS1; Thumpd3as1
Chromosome: 6
Strain of origin: C57BL/6NTac
Molecular note: A targeting vector was designed with (from 5' to 3') a ubiquitously expressed CAG promoter, a loxP-flanked stop cassette containing a neomycin resistance (neo) cassette, nuclear receptor binding SET domain protein 2 (Nsd2) cDNA, and a frt-flanked cassette containing an internal ribosome entry site (IRES) and an enhanced blue fluorescent protein (eBFP) sequence.