Human cDNA constructs
The wild-type version of human SMO (NM_601500) full length cDNA
clone was purchased from OriGene (SC122724, OriGene Technologies) and
subcloned into the mammalian expression vector pCS2+. Human SMOpoint mutation constructs were generated by PCR allowing for the
modification of each patient-associated single base alteration. All
human SMO point mutation variant constructs were generated using
Site-Directed Mutagenesis kit (QuickChange, Agilent Technologies)
according to the manufactures’ instructions without modification. All
subcloned cDNA constructs were verified by Sanger sequencing.