Primary Identifier | MGI:5008158 | Allele Type | Targeted |
Attribute String | Conditional ready, No functional change | Gene | Pi4ka |
Transmission | Germline | Strain of Origin | C57BL/6NTac |
Is Recombinase | false | Is Wild Type | false |
molecularNote | A targeting vector was designed to insert a loxP site upstream of exon 48 of the phosphatidylinositol 4-kinase, catalytic, alpha polypeptide (Pi4ka) gene. A human growth hormone polyadenylation signal (hGHpA) and a second loxP site followed by a frt/F3-flanked neomycin (neo) resistance cassette were placed downstream of exon 55 of Pi4ka. Using a recombination-mediated exchange cassette (RMCE), correctly targeted ES cells were then electroporated with a second targeting vector. This second targeting vector replaces the frt/F3-flanked neo cassette with a attB/attP-flanked puromycin (puro) resistance cassette and loxP site followed by a duplicated genomic region containing exons 48 to 55. The puro cassette and the duplicated region were flanked by frt/F3 sites. The duplicated genomic region contained a point mutation in exon 51 resulting in an Arginine to Lysine substitution (R1900K). PhiC31-mediated recombination removed the puro cassette. |