B6;129-Gt(ROSA)26Sor^{tm13(Nfkb-tdTomato)Nat}/J

Stock number: 037792
Product name: NFKB-LSL-nls-tdT-3xHA
Research areas: Research Tools

Description

NFKB-LSL-nls-tdT-3xHA reporter mice express nuclear-localized tdTomato (tdT) with a triple hemagglutinin (HA) epitope tag in response to NF-kappaB (NFKB) activation, but only in those cells that have a Cre recombinase-excised stop cassette.

Detailed description

NFKB-LSL-nls-tdT-3xHA reporter mice express nuclear-localized tdTomato (tdT) with a triple HA epitope tag in response to NF-kappaB (NFKB) activation, but only in those cells that have excised a loxP-transcription stop-loxP (LSL) cassette that is located between the promoter and the nls-tdT-3xHA open reading frame. Removing the LSL cassette in the germline generates a strain that can, in theory, respond to NFKB signaling in any cell. The tdTomato fluorescence may be faint, but anti-HA immunostaining can give a good clear nuclear signal.

In practice, the donating laboratory observed good reporter expression in vascular endothelial cells and in some immune cells in the context of experimental bacterial meningitis, but it is not known if the reporter response extends to other cell types.

Development

Five tandem NF-kappa-B (Nfkb) response elements (GGGAATTTCC; as found in Promega plasmid pGL4.32) were inserted upstream of a chicken minimal delta crystallin promoter (Crygd) followed by an intron. This is followed by a loxP-transcription stop-loxP (LSL) cassette, which is followed by a nuclear localized tdTomato (tdT) with a triple hemagglutinin (HA) epitope tag, which is followed by a bovine growth hormone (bGH) 3’ untranslated region. This entire cassette was targeted to the Gt(ROSA)26Sor locus in (129X1/SvJ x 129S1/Sv)F1-Kitl⁺-derived R1 embryonic stem (ES) cells by homologous recombination. An FRT-Neomycin-FRT selection cassette is retained immediately 5’ of the five tandem NF-kappa-B binding sites. This strain was maintained on a mixed C57BL/6 and 129 genetic background by the donating laboratory.

Upon arrival at The Jackson Laboratory, sperm was cryopreserved. Cryorecovery/live colony development will involve C57BL/6J (Stock No. 000664). (October, 2022)

Allele

Symbol: Gt(ROSA)26Sor^{tm13(Nfkb-tdTomato)Nat}
Name: targeted mutation 13, Jeremy Nathans
MGI accession ID: MGI:7365228
Generation method: Targeted
Attributes: Conditional ready (e.g. floxed); Reporter; Epitope tag
Synonyms: NFKB-LSL-nls-tdT-3xHA
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: (129X1/SvJ x 129S1/Sv)F1-Kitl⁺
Molecular note: Five tandem NF-kappa-B (Nfkb) response elements (GGGAATTTCC) are inserted upstream of a chicken minimal delta crystallin promoter (Crygd) followed by an intron. This is followed by a loxP-transcription stop-loxP (LSL) cassette, a nuclear localized tdTomato (tdT) with a triple hemagglutinin (HA) epitope tag, and a bovine growth hormone (bGH) in the 3' untranslated region. An FRT-Neomycin-FRT selection cassette is retained immediately 5'; of the five tandem NF-kappa-B binding sites.