RNA-seq analysis of mouse cardiomyocytes treated with dnMKL1

The goal of this study is to analyze the impact of dnMKL1 on transcription during cardiomyocyte maturation in mouse Overall design: Wildtype mice were injected with AAV9-dnMKL1-GFP or AAV9-GFP specifically in cardiomyocytes at P1. Five biological replicates were analyzed in each group. In each experiment, cardiomyocytes were isolated by retrograde collagenase perfusion and purified by fluorescence activated cell sorting. Total RNA was extracted from FACS sorted cells. RNA-seq libraries were generated using SMART-seq and Nextera kits and were sequenced on a NextSeq550 sequencer. FASTQ Data were aligned by STAR and count matrix generated by FeatureCounts.

本研究旨在分析显性负性MKL1（dnMKL1）对小鼠心肌细胞成熟过程中转录的影响。实验设计概述：野生型小鼠于出生后第1天（P1）特异性靶向心肌细胞注射AAV9-dnMKL1-GFP或AAV9-GFP载体。每组设置5个生物学重复样本。每次实验中，均采用逆行胶原酶灌流法分离心肌细胞，并通过荧光激活细胞分选（Fluorescence Activated Cell Sorting，FACS）进行纯化。从经荧光激活细胞分选纯化的细胞中提取总RNA。采用SMART-seq与Nextera建库试剂盒构建RNA测序文库，并在NextSeq550测序平台上完成测序。FASTQ数据通过STAR软件进行序列比对，通过FeatureCounts软件生成计数矩阵。