design: Expression of foetal gene Pontin is essential in protecting heart against pathological remodelling and cardiomyopathy
library_construction_protocol: Animal maintenance and experiments were conducted in accordance with the United Kingdom Animals (Scientific Procedures) Act and approved by the University of Manchester Ethics Committee. Pontin gene-targeted mice were obtained from the European Conditional Mouse Mutagenesis Programme (EUCOMM). The targeted allele was generated using a “knock-out first” strategy (PMID 21677750). The exon 3 of mouse Ruvbl1 gene (Pontin) was targeted using a construct carrying FRT and loxP sites. Heterozygous mice carrying the conditional targeted allele were crossed with CAG-Flpo transgenic mice to remove the lacZ and neo markers resulting in clean Pontin floxed (Pon flox) mice. To generate a cardiomyocyte specific inducible KO we crossed homozygous Pon flox mice with αMHC-MerCreMer transgenic mice (αMCM) generating αMCM- Pon flox mice. Pontin inducible cardiomyocyte knockout (Pontin icKO) were obtained by injecting 8-10 weeks old male αMCM- Pon flox mice with 40 mg/kg BW of tamoxifen. Heart tissues from PontinicKO mice (αMCM-Pon flox + TAM) and control (αMCM + TAM) were collected at 1 week and 3 week following tamoxifen injection.  RNA was extracted using Trizol reagent. Sequencing library for RNA sequencing was prepared with TruSeq RNA Library Prep Kit v2 (Illumina), according to the manufacturer's protocols.