A quantitative RT-PCR protocol to adapt and quantify RBM20-dependent exon splicing of targets at the human locus

Abstract

Summary: Gene splicing is a fine-tuned process orchestrated by splice factors including RNA-binding motif 20 (RBM20), and their mutations are linked to the development of cardiac diseases. Here, we provide a step-by-step protocol to transfer RBM20-dependent splicing from rat to human. This protocol describes a PCR-based approach to adapt and quantify RBM20-dependent exon-expression of human target genes. We detail the primer design, the use of induced pluripotent stem cell-derived cardiomyocytes (iPSC-CM) for RNA isolation, followed by quantification of splicing products.For complete details on the use and execution of this profile, please refer to Streckfuss-Bömeke et al. (2017).

Keywords

• Cell Biology
• Cell culture
• Gene Expression
• Molecular Biology
• Sequence analysis
• Sequencing