Gt(ROSA)26Sorem1(tetO-flpo*/ERT2,CAG-GFP*)Nijo
Endonuclease-mediated Allele Detail
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| Symbol: |
Gt(ROSA)26Sorem1(tetO-flpo*/ERT2,CAG-GFP*)Nijo |
| Name: |
gene trap ROSA 26, Philippe Soriano; endonuclease-mediated mutation 1, Nikhil Joshi |
| MGI ID: |
MGI:6383112 |
| Synonyms: |
NINJA |
| Gene: |
Gt(ROSA)26Sor Location: Chr6:113044389-113054205 bp, - strand Genetic Position: Chr6, 52.73 cM
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| Alliance: |
Gt(ROSA)26Sorem1(tetO-flpo*/ERT2,CAG-GFP*)Nijo page
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| Allele Type: |
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Endonuclease-mediated (Conditional ready, Epitope tag, Inducible, Recombinase, Reporter) |
| Inducer: |
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doxycycline/tetracycline |
| Mutation: |
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Insertion
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Gt(ROSA)26Sorem1(tetO-flpo*/ERT2,CAG-GFP*)Nijo expression driven by
1 gene
Knock-in expression driven by:
| Organism |
Driver Gene |
Note |
| bacteria |
tetO |
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Mutation details: The targeting vector is divided into two modules (regulatory and neoantigen) separated by a 2x chicken gamma globulin (CGG) insulator element. The regulatory module expresses an inducible flpo following cre recombination. The neoantigen module expresses GFP containing the peptide antigen, which is recognized by H2-Db-restricted CD8 T cells (GP-33-41) and H2-I-Ab restricted CD4 T cells (GP67-77) following flp recombination. The vector contains from 5' to 3': the regulatory module: loxP2-Puromycin resistance cassette - loxP1- C-terminal of flpo fused to human ERT2 (amino acids 251-596) in forward orientation separated by a pA and the N-terminal of flpo/ERT2 in the reverse orientation - loxP1 - loxP2 - tetO (pTRE tight promoter)-insulator element (blocks pCAG enhancer from activating pTRE), and the neoantigen module: CAG promoter - N terminal portion of GFP - FRT - FRT3 - neoantigen with a FLAG-TAG epitope - FRT - polyA - FRT3 - splice acceptor - premature STOP (alternative reading frame) - C terminal GFP - polyA. The neoantigen is comprised of sequence from the lymphocytic meningitis virus (LCMV) glycoprotein modified such that the DNA sequence is split across three exons with exon 2 inverted and flanked by non-compatible FRT sites (F3 and FRT). The targeting vector was inserted into the Rosa26 locus. In the presence of flpo, exon 2 is inverted lining up splicing sites to allow transcription of the full-length GFP containing the peptide antigen.
(J:317156)
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Activity: |
 Tissue activity of this recombinase allele
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Driver:
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tetO
(bacteria)
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| Original: |
J:317156 Damo M, et al., Inducible de novo expression of neoantigens in tumor cells and mice. Nat Biotechnol. 2020 Jul 27; |
| All: |
3 reference(s) |
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Analysis Tools
