Novel heterozygous mutations of the SF1 gene and their functional characterization in patients with 46,XY disorders of sex development (DSD) without adrenal insufficiency

Methods This study included 48 patients with 46,XY DSD without adrenal insufficiency or dysmorphic features. Genomic DNA was extracted from peripheral blood leukocytes and direct sequencing of the 6 coding exons of SF1 was performed. To evaluate the effect of SF1 mutations on transcriptional activity, transient transfection studies were performed using dual luciferase reporter assay system with cotransfection of PGL4.75 Renilla luciferase as a marker of transfection efficiency. Wild-type, or mutant SF1 expression vectors with SF1 dependent StAR and MIS promoters linked to luciferase were assayed for luciferase activity.