Gene silencing using adenoviral RNAi vector in vascular smooth muscle cells and cardiomyocytes.
RNA interference (RNAi) is a new and rapidly progressing technology for facilitating functional gene silencing. To perform highly efficient RNAi in cardiomyocytes, vascular smooth muscle cells, and vascular endothelial cells, which are known to have very low transfection efficiency, adenovirus-mediated RNAi was employed. The effects of RNAi on GAPDH transcripts were successfully reduced by nearly 90% in the primary cultured cells, indicating that adenovirus-mediated gene silencing is a promising technique for gene silencing in cardiovascular studies. This chapter describes general guidelines for selecting RNAi target sites, construction of a shuttle vector encoding short hairpin RNAi, and generation of recombinant adenovirus.
Kasahara
Hideko
H
Department of Physiology and Functional Genomics, University of Florida College of Medicine, Gainesville, FL, USA.
Aoki
Hiroki
H
D000256:Adenoviridae / Q000235:genetics; D000818:Animals; D003001:Cloning, Molecular; D020868:Gene Silencing; D005822:Genetic Vectors; D006801:Humans; D009131:Muscle, Smooth, Vascular / Q000166:cytology / Q000378:metabolism*; D032383:Myocytes, Cardiac / Q000166:cytology / Q000378:metabolism*; D011401:Promoter Regions, Genetic; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000235:genetics / Q000378:metabolism; D014158:Transcription, Genetic
D034741:RNA, Small Interfering