GSE132992
CRISPR-mediated activation of endogenous gene expression in the postnatal heart [RNA-seq]
Endogenous activation of Myocyte enhancer factor 2 D (Mef2d) in the postnatal mouse heart by cardiomyocyte-specific CRISPRactivation (Myh6-dCas9VPR) The aim of this study was to compare endogenous gene activation of the pro-hypertrophic factor Myocyte enhancer factor 2 D (Mef2d) in the postnatal mammalian heart and experimental control groups including wild-type litter mates injected with saline, wild-type litter mates injected with AAV9 TRISPR Mef2d and transgenic litter mates injected with saline. Mice were injected with saline/AAV9 TRISPR Mef2d at P4 and sacrificed 8 weeks later. RNA was isolated by TRIzol/Chloroform and isopropanol precipitation. Ventricular tissue mRNA profiles were obtained using deep sequencing, in triplicates, using Illumina HiSeq4000. The sequence reads that passed quality filters were analyzed at the transcript isoform level with TopHat followed by DESeq2. qPCR validation was performed using SYBR Green assays. We present in this study a mouse model for cardiomyocyte-specific endogenous gene activation in the postnatal mammalian heart by CRISPR/dCas9VPR. As a proof of concept, we activated the pro-hypertrophic factor Myocyte enhancer factor 2 D (Mef2d) leading to functional decline, increased marker expression of heart failure as well as cardiomyocyte hypertrophy over a time course of 8 weeks. We furthermore observed no phenotypic consequence of constitutive dCas9VPR transgene expression in the postnatal heart. We therefore conclude, that the Myh6-dCas9VPR mouse model is a suitable tool for endogenous gene activation studies in the postnatal heart. Overall design: Transcriptome profiling from Mef2d endogenous gene activation in the postnatal hearts versus controls.