Small interfering RNA-induced transcriptional gene silencing in human cells.
Small interfering RNA (siRNA) and microRNA silence genes at the transcriptional, posttranscriptional, and/or translational level. Using human tissue culture cells, we show that promoter-directed siRNA inhibits transcription of an integrated, proviral, elongation factor 1alpha (EF1A) promoter-green fluorescent protein reporter gene and of endogenous EF1A. Silencing was associated with DNA methylation of the targeted sequence, and it required either active transport of siRNA into the nucleus or permeabilization of the nuclear envelope by lentiviral transduction. These results demonstrate that siRNA-directed transcriptional silencing is conserved in mammals, providing a means to inhibit mammalian gene function.
Department of Medicine 0678, Stein Clinical Research Building, Room 302, University of California-San Diego, La Jolla, CA 92093-0678, USA. kmorris@coh.org
Kevin V
KV
Morris
Simon W-L
SW
Chan
Steven E
SE
Jacobsen
David J
DJ
Looney
D021581:Active Transport, Cell Nucleus; D002460:Cell Line; D002467:Cell Nucleus / Q000378:metabolism; D019175:DNA Methylation; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism; D015536:Down-Regulation; D015977:Eukaryotic Initiation Factor-1 / Q000235:genetics*; D020868:Gene Silencing; D017930:Genes, Reporter; D005822:Genetic Vectors; D049452:Green Fluorescent Proteins; D006801:Humans; D016089:Immunodeficiency Virus, Feline / Q000235:genetics; D008164:Luminescent Proteins / Q000096:biosynthesis / Q000235:genetics; D011401:Promoter Regions, Genetic; D034741:RNA, Small Interfering / Q000235:genetics / Q000378:metabolism*; D011993:Recombinant Fusion Proteins; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014158:Transcription, Genetic; D014161:Transduction, Genetic; D014162:Transfection; D019076:Transgenes
D004268:DNA-Binding Proteins; D015977:Eukaryotic Initiation Factor-1; D008164:Luminescent Proteins; C475074:MPG peptide; D034741:RNA, Small Interfering; D011993:Recombinant Fusion Proteins; C026022:eukaryotic peptide initiation factor-1A; D049452:Green Fluorescent Proteins