Gene silencing of Col1α1 by RNAi in rat myocardium fibroblasts

Abstract

Abstract Objective The collagen type I alpha 1 (Col1α1) not only acts as a scaffold for the cell matrix, but also is involved in myocardial fibrosis. The purpose of this study is to screen an efficient shRNA specific to rat Col1α1 gene. Methods The shRNA expression plasmids targeting Col1α1 were constructed and their knockdown efficiency was examined by qPCR and Western blot. Results Three shRNA expression plasmids targeting Col1α1 were successfully constructed, as suggested by analysis of restriction enzyme digestion and DNA sequencing. The Col1α1 shRNA-expressing plasmids were then transfected into rat cardiac fibroblasts with high efficiency. The qPCR and Western blotting results demonstrated that Col1α1 expression at both mRNA and protein level was significantly inhibited (p < 0.05) in shRNA-transfected cells compared to scrambled shRNA-transfected cells. Conclusion A Col1α1-specific shRNA with high efficiency of gene silencing is successfully obtained. The construction of Col1α1 shRNA-expressing plasmid lays the foundation for the further studies on the role of Col1α1 gene in myocardial fibrosis.