C57BL/6-Gt(ROSA)26Sor^{tm6(Map3k14*)Rsky}/J

Stock number: 012638
Research areas: Immunology, Inflammation and Autoimmunity Research, Research Tools

Description

Mice homozygous for the NIK-deltaT3 allele are viable and fertile, with a loxP-flanked Neo-STOP cassette preventing transcription of the downstream bicistronic sequences (NIKdeltaT3 [a constitutively active form of the mouse NF-kappaB inducing kinase] and EGFP).

Detailed description

Mice homozygous for the NIK-deltaT3 allele are viable and fertile, with a loxP-flanked Neo-STOP cassette preventing transcription of the downstream bicistronic sequences (NIKdeltaT3 [a constitutively active form of the mouse NF-kappaB inducing kinase] and EGFP). When bred to mice that express Cre recombinase, offspring will have the STOP cassette deleted and subsequent expression of the NIKdeltaT3 molecule and EGFP fluorescence in the cre-expressing cells. Expression of NIK-deltaT3 leads to uncoupling NIK from tumor necrosis factor receptor-associated factor 3 (TRAF3)-mediated control causing maximal p100 processing and dramatic hyperplasia of B cell-activating factor of the TNF family receptor (BAFF-R)-independent B cells. BAFF-R binding is required to sustain the NIK p100 subunit processing needed for B cell survival, as implicated in human multiple myeloma.

Of note, breeding these mice to an FLP-expressing strain will result in removal of the frt-flanked IRES-EGFP cassette.

When bred to a strain expressing Cre recombinase throughout B lymphocyte development and differentiation (see Stock No. 006785 for example), this mutant mouse strain exhibits increased B cell numbers.

Development

The NIK-deltaT3 targeting vector was designed with (from 5' to 3') a loxP-flanked neomycin resistance-STOP cassette, a cDNA encoding a FLAG-tagged, constitutively active form of murine NF-kappaB inducing kinase (NIKdeltaT3, NIK mutant lacking the Traf3 binding domain), a frt-flanked internal ribosomal entry site and enhanced green fluorescent protein (IRES-EGFP), and a bovine polyA sequence. This construct was placed under control of the promoter of the targeted gene via electroporation into C57BL/6-derived Bruce-4 embryonic stem (ES) cells. Correctly targeted ES cells were selected, injected into blastocysts and chimeric animals were bred to C57BL/6 mice to generate mutant mice. These R26-STOP-NIKdelta3 mice were maintained as a homozygous colony for many generations. Upon arrival at The Jackson Laboratory, mutant mice were bred to C57BL/6J inbred mice (Stock No. 000664) for at least one generation to establish the colony.

Allele

Symbol: Gt(ROSA)26Sor^{tm6(Map3k14*)Rsky}
Name: targeted mutation 6, Klaus Rajewsky
MGI accession ID: MGI:3809723
Generation method: Targeted
Attributes: Conditional ready (e.g. floxed); Reporter; Inserted expressed sequence
Marker symbol: Gt(ROSA)26Sor
Marker name: gene trap ROSA 26, Philippe Soriano
Chromosome: 6
Strain of origin: B6.Cg-Thy1^a
Expressed genes: Map3k14,mitogen-activated protein kinase kinase kinase 14,mouse, laboratory; GFP,Green Fluorescent Protein,
Site of expression: When bred to mice that express Cre recombinase, bicistronic expression of GFP and NIKdeltaT3 (a constitutively active form of the mouse NF-kappaB inducing kinase, Map3k14) will occur in cells where promoters driving Cre recombinase are expressed.
Molecular note: The gene lacking the sequence that encodes the TRAF3-binding domain, preceded by a floxed neo STOP cassette and followed by an frt-flanked IRES-EGFP cassette, was knocked into the locus to achieve cre-inducible, ubiquitous expression. Cre mediated removal of the STOP cassette is required for expression.