Cloning of the Mutant hTopIIIα Expression Vector
A plasmid encoding the wild-type (WT) TOP3A cDNA24 was modified by the Quickchange XL Site-Directed Mutagenesis Kit (Agilent technologies) with the following primers to recapitulate the deletion and frameshift present in subject P1: 5′-CCCTCCGTCACACGACTGTGCAGAAGGA-3′ (T3_FS_FW) and 5′-TCCTTCTGCACAGTCGTGTGACGGAGGG-3′ (T3_FS_RW).