Retrovirus vector-mediated stable gene silencing in human cell.
RNA interference (RNAi) is the mechanism of sequence-specific, post-transcriptional gene silencing initiated by short interfering RNAs (siRNAs) homologous to the gene being suppressed. siRNAs, which mediate sequence-specific mRNA degradation, are duplexes of about 21-23 nucleotides with 3'-overhangs synthesized in vitro or expressed by DNA-based vector. However, these systems rely on transfection for delivery and cannot generate long-term gene silencing in vivo. This obstacle may be circumvented by recently developed retrovirus- and lentivirus-delivered RNAi. Here, we describe a retroviral system for delivery of siRNA into cells, which can substantially down-regulate the expression of human p53 gene in human HepG2 cells. What's more, the G1 and S phases of cell cycle change dramatically in p53-down-regulated cells. These results indicate that retrovirus vector-delivered RNAi may be used in functional genomics and in gene therapy.
Liu
Chang-Mei
CM
National Laboratory of Medical Molecular Biology, Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences and Peking Union Medical College, 5 Dong Dan San Tiao, Beijing 100005, PR China.
Liu
De-Pei
DP
Dong
Wen-Ji
WJ
Liang
Chih-Chuan
CC
D001483:Base Sequence; D015153:Blotting, Western; D002460:Cell Line; D004247:DNA / Q000378:metabolism; D015536:Down-Regulation; D020868:Gene Silencing; D016158:Genes, p53; D005822:Genetic Vectors; D005838:Genotype; D006801:Humans; D008969:Molecular Sequence Data; D010641:Phenotype; D016133:Polymerase Chain Reaction; D034622:RNA Interference; D012333:RNA, Messenger / Q000378:metabolism; D034741:RNA, Small Interfering / Q000378:metabolism; D012190:Retroviridae / Q000235:genetics*; D014162:Transfection
D012333:RNA, Messenger; D034741:RNA, Small Interfering; D004247:DNA