Neeraj Sharma, Patrick R Sosnay, Anabela S Ramalho, Christopher Douville, Arianna Franca, Laura B Gottschalk, Jeenah Park, Melissa Lee, Briana Vecchio-Pagan, Karen S Raraigh, Margarida D Amaral, Rachel Karchin, Garry R Cutting
Assessment of the functional consequences of variants near splice sites is a major challenge in the diagnostic laboratory. To address this issue, we created expression minigenes (EMGs) to determine the RNA and protein products generated by splice site variants (n = 10) implicated in cystic fibrosis (CF). Experimental results were compared with the splicing predictions of eight in silico tools. EMGs containing the full-length Cystic Fibrosis Transmembrane Conductance Regulator (CFTR) coding sequence and flanking intron sequences generated wild-type transcript and fully processed protein in Human Embryonic Kidney (HEK293) and CF bronchial epithelial (CFBE41o-) cells...
October 2014: Human Mutation