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

ABSTRACT: 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).

SUBMITTER: Rebs S

PROVIDER: S-EPMC8784397 | biostudies-literature | 2022 Mar

REPOSITORIES: biostudies-literature

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

Rebs Sabine S Buchwald Tjark Alexander TA Streckfuss-Bömeke Katrin K

STAR protocols 20220119 1

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 ...[more]

PMID: 35106501

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