TgTn(pb-Gt(ROSA)26Sor/CAG-EYFP*,-mCherry*,-Notch1*,-EGFP*,-Rbpj*,-mCerulean*)#Ben
Transgene Detail

Summary

• Symbol: TgTn(pb-Gt(ROSA)26Sor/CAG-EYFP*,-mCherry*,-Notch1*,-EGFP*,-Rbpj*,-mCerulean*)#Ben
• Name: transgenic transposon concatemer, Rui Benedito
• MGI ID: MGI:6761333
• Synonyms: iChr-Notch-v2-Mosaic, TgBAC(Rosa)26Sor^{(iChr-Notch-v2-Mosaic)}
• Transgene: TgTn(pb-Gt(ROSA)26Sor/CAG-EYFP*,-mCherry*,-Notch1*,-EGFP*,-Rbpj*,-mCerulean*)#Ben Location: unknown
• Alliance: TgTn(pb-Gt(ROSA)26Sor/CAG-EYFP*,-mCherry*,-Notch1*,-EGFP*,-Rbpj*,-mCerulean*)#Ben page

Transgene origin

• Germline Transmission: Earliest citation of germline transmission: J:252386
• Parent Cell Line: G4 (ES Cell)
• Strain of Origin: (129S6/SvEvTac x C57BL/6NCrl)F1

Transgene description

• Transgene Type: Transgenic (Conditional ready, Epitope tag, Inserted expressed sequence, Reporter, Transposon concatemer)
• Mutation: Transposon insertion
• TgTn(pb-Gt(ROSA)26Sor/CAG-EYFP*,-mCherry*,-Notch1*,-EGFP*,-Rbpj*,-mCerulean*)#Ben expresses 2 genes
  Transgene expresses:
  

  Organism	Expressed Gene	Note
  mouse	Notch1 (MGI:97363)	sequence coding for the intra-cellular and PEST domains is downstream in-frame with T2A sequence which is downstream in-frame with mCherry gene
  mouse	Rbpj (MGI:96522)	sequence coding for mutated mouse peptide with p.R218H substitution is downstream in-frame with T2A sequence which is downstream in-frame with EGFP gene

• Mutation details: This allele has three chromatin/nuclear-targeted fluorescent protein sequences uniquely flanked with pairs of incompatible lox sites (in the same direction), all inserted between exons 1 and 2 of the Gt(ROSA)26Sor locus on BAC RP23-401D9. PiggyBAC transposon inverted repeats were added to the BAC to facilitate single-copy and complete insertion into the genome with the PiggyBAC system. The targeting vector is designed with (from 5' to 3') two copies of chicken beta-globin HS4 insulator element (2xINS; increases downstream gene expression and minimizes regulatory interference), a CMV-IE enhancer/chicken beta-actin/rabbit beta-globin hybrid promoter (from pCAGGS), a loxN site, a lox2272 site, a loxP site, the N-PhiMut gene (non-cytotoxic/non-fluorescent Phialidium yellow fluorescent protein variant PhiYFP with nuclear localization signal), bovine GH polyA signal sequence, a second loxN site, the HA-H2B-Cherry gene (monomeric DsRed fluorescent protein variant mCherry with N-terminal hemagglutinin (HA) tag and 125 amino acid human histone 2B chromatin/nucleus tag sequence), in-frame Thosea asigna virus 2A cleavage site (T2A; mediates ribosomal skipping), in-frame mouse Notch1 intracellular domain (NICD) and PEST domain sequence (NICD-PEST), bovine GH polyA signal sequence, a second lox2272 site, the H2B-EGFP-V5 gene (enhanced green fluorescent protein with 127 amino acid and human histone 2B chromatin/nucleus tag plus C-terminal paramyxovirus V5 tag sequence), in-frame T2A sequence, bovine GH polyA signal sequence, a second loxP site, the His-H2B-Cerulean gene (monomeric enhanced cyan fluorescent protein variant mCerulean with N-terminal 6xHis affinity tag and 125 amino acid human histone 2B chromatin/nucleus tag sequence), in-frame T2A sequence, in-frame sequence for a dominant-negative form of mouse Rbpj (DN-Rbpj, with a p.R218H mutation), bovine GH polyA signal sequence, an FRT site flanked neomycin resistance gene cassette, and a second 2xINS sequence. The neo cassette was removed through subsequent flp-mediated recombination. (J:252386)

Find Mice (IMSR)

• Mouse strains and cell lines available from the International Mouse Strain Resource (IMSR)
• Carrying this Mutation: Mouse Strains: 0 strains available Cell Lines: 0 lines available

References

• Original: J:252386 Pontes-Quero S, et al., Dual ifgMosaic: A Versatile Method for Multispectral and Combinatorial Mosaic Gene-Function Analysis. Cell. 2017 Aug 10;170(4):800-814.e18
• All: 2 reference(s)