B6;129S6-Gt(ROSA)26Sortm2(CAG-NuTRAP)Evdr/J

Cat. No.: CEMM-07250767

Common Name: NuTRAP; RANGAP-TRAP
The NuTRAP allele (Nuclear tagging and Translating Ribosome Affinity Purification) has a loxP-flanked STOP preventing transcription of three individual components: BirA, BLRP-tagged mCherry/mRANGAP1 and EGFP/L10a. When expressed, the BLRP-tagged mCherry/mRANGAP1 protein is biotinylated by BirA, allowing nuclear membrane-labeling with mCherry and biotin (which enables nuclear isolation by fluorescence- and affinity-based purification). Furthermore, EGFP/L10a fluorescently tags the translating mRNA polysome complex (which enables isolation of RNAs that are actively engaged by ribosomes). Overall, NuTRAP mice are a Cre-inducible tool strain that allows labeling and simultaneous isolation of cell type-specific nuclei and mRNA, and are well-suited for studying epigenomics and transcriptomics from specific cell types within a heterogeneous tissue.
Inquiry
Status Live Mouse
Frozen Embryo
Age 4 weeks
12 weeks
Customized Age
Sex Male
Female
GENETICS
Allele Symbol
Gt(ROSA)26Sortm2(CAG-NuTRAP)Evdr
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Allele Name
targeted mutation 2
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Allele Attributes
Conditional ready (e.g. floxed); Reporter
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Gene Symbol
Gt(ROSA)26Sor
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Gene Name
gene trap ROSA 26
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Chromosome
6
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Expressed Genes
RFP, Red Fluorescent Protein, coral; GFP, Green Fluorescent Protein
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MGI Accession ID show more close
Site of Expression
Upon exposure to Cre recombinase, the NuTRAP allele co-expresses BirA (E. Coli biotin ligase), BLRP (biotin ligase recognition peptide)-tagged mCherry/mRANGAP1 (the mouse nuclear membrane RAN GTPase activating protein) and EGFP/L10a (the 60S ribosomal subunit L10a) fusion proteins in Cre expressing tissues.
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Strain of Origin
(129S6/SvEvTac x C57BL/6NCrl)F1
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Molecular Note
The targeting vector was designed with (from 5' to 3') with a CMV-IE enhancer/chicken beta-actin/rabbit beta-globin hybrid promoter, a FRT site, a loxP-flanked STOP cassette, the E. Coli biotin ligase gene (birA), a 2A peptide, the BLRP-mCherry-mRANGAP1 fusion gene (described below), a 2A peptide, the EGFP-L10a fusion protein (an enhanced green fluorescent protein gene [EGFP] fused in-frame to the N-terminus of a cDNA sequence encoding the mouse 60S ribosomal subunit L10a [Rpl10a]), a WPRE, a polyA signal, an AttB site, a PGK-FRT Neo-PGK polyA, and an AttP site. This entire targeting vector was inserted between exons 1 and 2 of the Gt(ROSA)26Sor locus. The BLRP-mCherry-mRANGAP1 fusion gene consists of a monomeric red fluorescent protein sequence fused in-frame to the N-terminal of a cDNA sequence encoding the mouse nuclear membrane RAN GTPase activating protein 1. This fusion gene is tagged at the mCherry N-terminal with a biotin ligase recognition peptide (BLRP ; a 23 amino acid protein with AviTag core.
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HUSBANDRY
Suggested Controls
C57BL/6J
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Breeding Considerations
When maintaining a live colony, heterozygous mice may be bred together, to wildtype mice from the colony or to C57BL/6J inbred mice. Alternatively, homozygous mice may be bred together.
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Breeding Strategy
Homozygote x Homozygote
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For Research Use Only.
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