Effect of cardiomyocyte KLF5 deletion in cardiac gene expression profile

Kruppel-like factor 5 gene was deleted (Cre-lox) system specifically in cardiomyocytes (aMHC-promoter-driven expression of Cre) and RNA was purified from total hearts (TRIZOL method) following perfusion of the heart with PBS. Basal levels cardiac RNA was analyzed with whole genome microarray chips. There were two groups of mice: 4 mice that had KLF5 gene floxed without expressing Cre recombinase (control group) and 4 mice that were expressing Cre in cardiomyocytes and therefore KLF5 gene was deleted and thus not expressed in this particular cell type.本实验采用Cre-lox重组酶系统，在心肌细胞中特异性敲除Krüppel样因子5（Kruppel-like factor 5, KLF5）基因，其中Cre重组酶的表达由aMHC启动子驱动。经磷酸盐缓冲液（PBS）灌流心脏后，采用TRIZOL法从完整心脏组织中提取总RNA。对基础状态下的心肌总RNA使用全基因组微阵列芯片进行分析。实验分为两组小鼠：对照组为4只携带KLF5基因floxed等位基因但不表达Cre重组酶的小鼠；实验组为4只在心肌细胞中表达Cre重组酶的小鼠，其心肌细胞内KLF5基因已被敲除，因此该细胞类型中无KLF5基因表达。