Applications of gene transfer to study rna splicing in mammalian cell lines

Summary The first methods available for studying the signals required for splicing of eukaryotic pre-mRNA involved the transfection of cloned genes into cells in culture. The method is still of considerable importance for investigating splice site selection and for identifying the signals that modulate alternative patterns of splicing during differentiation. A method is described for the transfection of myoblast cell lines. The techniques available for preparation of RNA from transfected cells are reviewed and procedures are given for the analysis of splice site use by mapping with nuclease S1 or RT-PCR.