RBPMS and RBPMS2 Cooperate to Safeguard Cardiac Splicing E11.5

RBPMS and RBPMS2 share a high amino acid sequence similarity and are interchangeable in in vitro splicing assays. However, it remains unclear whether RBPMS and RBPMS2 play similar or distinct function during heart development in vivo. To address this question, we constructed Rbpms and Rbpms2 floxed mice with an established CRISPR-Cas9 method, and validated the efficacy of Rbpms or Rbpms2 floxed alleles in ablating RBPMS or RBPMS2 proteins by crossing them with germline-expressing Sox2-Cre. Transcriptome were abtained by RNA seq to investigate the cardiac function of RBPMS and RBPMS2. We performed RNA sequencing on RbpmscmKO, Rbpms2cmKO, and Rbpms/Rbpms2dcmKO hearts along with their littermate controls. In this study, we generated cardiomyocyte-specific knockout (KO) mice for Rbpms, Rbpms2, and both genes combined (double KO). The experimental groups included Rbpms single KO, Rbpms2 single KO, and Rbpms/Rbpms2 double KO mice, which were analyzed to assess cardiac phenotypes. RNA sequencing was conducted on E11.5 heart tissue from each KO group to evaluate changes in gene expression and splicing patterns. In silico analyses were used to dissect the mechanisms behind the distinct and overlapping roles of RBPMS and RBPMS2 in splicing regulation during heart development. The study focused on identifying the effects of RBPMS and RBPMS2 on sarcomere assembly and the regulation of cardiac-specific splicing programs, as well as determining the impact of their binding locations on pre-mRNA.