Common haplotypes for each of 14 promoter and UTR regions were PCR-amplified and cloned into luciferase-reporter vectors. Promoter haplotypes were cloned immediately upstream of the luciferase reporter gene, while 3′UTRs were placed at the 3′ end of a luciferase gene whose expression is driven by the RPL10 promoter that has strong constitutive activity (vector maps available from SwitchGear Genomics, http://switchgeargenomics.com/resources/vector-maps/). We then transfected each of these constructs into HEPG2 cells, a liver-derived cell line, and measured luminescence. Each haplotype was tested using multiple (mode = 3) vector preparations and 4 technical transfection replicates measurements were obtained for each vector preparation (12 or more measurements for most haplotypes).
