Gt(ROSA)26Sor^{em1(tetO-flpo*/ERT2,CAG-GFP*)Nijo}
Endonuclease-mediated Allele Detail

Summary

• Symbol: Gt(ROSA)26Sor^{em1(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
• Alliance: Gt(ROSA)26Sor^{em1(tetO-flpo*/ERT2,CAG-GFP*)Nijo} page

Mutation origin

• Strain of Origin: C57BL/6N

Mutation description

• Allele Type: Endonuclease-mediated (Conditional ready, Epitope tag, Inducible, Recombinase, Reporter)
• Inducer: doxycycline/tetracycline
• Mutation: Insertion
• Gt(ROSA)26Sor^{em1(tetO-flpo*/ERT2,CAG-GFP*)Nijo} expression driven by 1 gene
  Knock-in expression driven by:
  

  Organism	Driver Gene	Note
  bacteria	tetO

• 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)

Recombinase activity

Activity:	Tissue activity of this recombinase allele
Driver:	tetO (bacteria) Summary of all recombinase alleles driven by tetO.

Find Mice (IMSR)

• Mouse strains and cell lines available from the International Mouse Strain Resource (IMSR)
• Carrying this Mutation: Mouse Strains: 1 strain available Cell Lines: 0 lines available
• Carrying any Gt(ROSA)26Sor Mutation: 993 strains or lines available

References

• 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)