Inhibiting gene expression at transcription start sites in chromosomal DNA with antigene RNAs.
Transcription start sites are critical switches for converting recognition of chromosomal DNA into active synthesis of RNA. Their functional importance suggests that they may be ideal targets for regulating gene expression. Here, we report potent inhibition of gene expression by antigene RNAs (agRNAs) complementary to transcription start sites within human chromosomal DNA. Silencing does not require methylation of DNA and differs from all known mechanisms for inhibiting transcription. agRNAs overlap DNA sequences within the open complex formed by RNA polymerase, and silencing is acutely sensitive to single base shifts. agRNAs effectively silence both TATA-less and TATA-box-containing promoters. Transcription start sites occur within every gene, providing predictable targets for agRNAs. Potent inhibition of multiple genes suggests that agRNAs may represent a natural mechanism for controlling transcription, may complement siRNAs and miRNAs that target mRNA, and will be valuable agents for silencing gene expression.
Janowski
Bethany A
BA
Department of Pharmacology, University of Texas Southwestern Medical Center at Dallas, Dallas, Texas 75390, USA.
Huffman
Kenneth E
KE
Schwartz
Jacob C
JC
Ram
Rosalyn
R
Hardy
Daniel
D
Shames
David S
DS
Minna
John D
JD
Corey
David R
DR
D001483:Base Sequence; D002478:Cells, Cultured; D002877:Chromosomes, Human; D004247:DNA / Q000037:antagonists & inhibitors* / Q000378:metabolism*; D005786:Gene Expression Regulation / Q000187:drug effects; D020868:Gene Silencing; D006801:Humans; D008969:Molecular Sequence Data; D034741:RNA, Small Interfering / Q000378:metabolism* / Q000494:pharmacology; D024363:Transcription Initiation Site / Q000187:drug effects / Q000502:physiology*
D034741:RNA, Small Interfering; D004247:DNA