Primary Identifier | MGI:7714771 | Allele Type | Targeted |
Attribute String | Not Specified | Gene | Rr350 |
Transmission | Germline | Strain of Origin | C57BL/6 |
Is Recombinase | false | Is Wild Type | false |
molecularNote | Through recombinase-mediated genomic replacement (RMGR) (a type of recombinase-mediated cassette exchange, RMCE) using ESCs containing the Rhbdf1tm1Ajhs and Hbatm1Paz alleles and where the Hba cluster was deleted, the endogenous sequence was replaced with a synthetic BAC where hematopoietic Hba enhancers R1 (Rr349), R3 (Rr352) and R4 (Rr350) are intact and all endogenous sequences between the Rhbdf1 and Hba allele endpoints are present. The targeted sequence has an FRT site, 5' Hprt sequences, loxP site, neomycin resistance gene cassette and a second loxP site in an intron of Rhbdf1 and a puromycin resistance gene cassette, tk gene and lox511 site in an intron of Hbq1a. The replacement BAC has a loxP site, hygromycin resistance gene cassette, a second loxP site, 3' Hprt sequences and an FRT site in the same intron of Rhbdf1 and a lox511 site, neomycin resistance gene cassette and tk gene in the same intron of Hbq1a. After Cre-mediated sequence exchange, the modified genome contains an FRT site flanked reconstituted Hprt minigene (with a loxP site in an intron) and a lox511 site in Rhbdf1 and Hbq1a, respectively. The Hprt gene was removed through subsequent Flp-mediated recombination leaving one FRT and one lox511 site. This allele essentially represents a reconstituted wild-type allele. |