An arrhythmogenic cardiomyopathy pig model by PKP2 heterozygous knockout

Patients with arrhythmogenic cardiomyopathy (ACM) have a high incidence of ventricular arrhythmias and a significant risk of sudden death. However, the difficulty in obtaining myocardial tissue samples from ACM patients at the early stage of the disease, coupled with the lack of animal models that can simulate the characteristics of human ACM, has severely hindered in-depth research into its pathogenesis. Overall design: Total RNA was extracted from right ventricular tissue of PKP+/- and WT pig using RNeasy Mini Kit.). A total amount of 1 µg RNA per sample was used as input material for the RNA sample preparations. Sequencing libraries were generated using NEBNext® UltraTM RNA Library Prep Kit for Illumina® (NEB, USA) following manufacturer's recommendations and index codes were added to attribute sequences to each sample. The library preparations were sequenced on an Illumina Novaseq platform and 150 bp paired-end reads were generated. The index of the reference genome was built using Hisat2 v2.0.5. and paired-end clean reads were aligned to the reference genome using Hisat2 v2.0.5.

致心律失常性心肌病（arrhythmogenic cardiomyopathy, ACM）患者的室性心律失常发生率较高，且猝死风险显著。但由于疾病早期难以获取ACM患者的心肌组织样本，加之缺乏可模拟人类ACM特征的动物模型，严重阻碍了对其发病机制的深入研究。整体实验设计：使用RNeasy Mini Kit从PKP+/-与野生型（wild type, WT）猪的右心室组织中提取总RNA。每个样本取1 µg总RNA作为起始材料开展RNA样品制备。按照制造商操作指南，使用NEBNext® UltraTM RNA文库制备试剂盒（适配Illumina®平台，NEB，美国）构建测序文库，并添加索引编码以区分各样本的测序序列。构建完成的文库在Illumina Novaseq平台进行测序，生成150 bp双端读段。使用Hisat2 v2.0.5构建参考基因组索引，并通过该软件将双端清洁读段比对至参考基因组。