Tc(HSA19*)SDkn

The allele is created in a multi-step process by exchanging approx. 20 kb on mouse chromosome 7 stretching from and including Tomm40 to Apoe from BAC RP24-136M6 with an acceptor cassette containing PGK-hygromycin and HSV-TK flanked by loxP/lox2272 sites using C57BL/6N-derived C2(Nagy) embryonic stem cells (ES). The 5' loxP site is located downstream of Pvrl2 and the 3'; lox2272 site is located downstream of the last Apoe exon (including the enhancer segment). ES cells carrying the acceptor cassette were electroporated with the exchange vectors and a plasmid carrying Cre recombinase to insert 23.77 kbp from human BAC (RP11-84C16) containing TOMM40 to APOE and the flanking sequences (including promoter and regulatory regions) followed by an FRT-flanked PGK-neo cassette and a loxP site on chr19 : 45,393,394-45,417,163 (HSA19). The modified BAC carries the APOE episilon 3 backbone and the TOMM40 523 polyT short allele (S, T=15). APOE epsilon4 is converted to APOE epsilon3 by exchanging the APOE epsilon4 arginine (CGC) at SNP rs429358 with the APOE epsilon3 cysteine (TGC). (J:341425)