Primary Identifier | MGI:7859502 | Allele Type | Transposon induced |
Attribute String | Reporter | Gene | Tn(Nr4a1-mCherry*)M4Cya |
Strain of Origin | C57BL/6J | Is Recombinase | false |
Is Wild Type | false |
molecularNote | Bacterial artificial chromosome (BAC) RP24-366J14 containing the mouse gene Nr4a1 and regulatory regions was used as the starting template. A Fast-FT-rBG-pA-Zeomycin selection cassette was inserted into exon 2 (first coding exon) of Nr4a1 via homologous recombination. The Zeomycin cassette was subsequently removed by homologous combination. PiggyBac ITR-flanked ampicillin selection cassette was introduced into the BAC vector backbone and the SacBII and loxP sites were replaced. The two ITR elements are recognition sequences of the piggyBac transposase (PBase). Amp expression cassettes were used for selection of correctly modified BACs. The modified BAC was injected into C57BL/6J single-cell stage fertilized eggs. The region from 5â ITR to 3â ITR on the BAC (which contains the Nr4a1-FT-Fast allele) were transposed into random TTAA sites of the host genome mediated by PBase, with one integration copy per site. |