Exogenous control of mammalian gene expression through modulation of RNA self-cleavage.
Recent studies on the control of specific metabolic pathways in bacteria have documented the existence of entirely RNA-based mechanisms for controlling gene expression. These mechanisms involve the modulation of translation, transcription termination or RNA self-cleavage through the direct interaction of specific intracellular metabolites and RNA sequences. Here we show that an analogous RNA-based gene regulation system can effectively be designed for mammalian cells via the incorporation of sequences encoding self-cleaving RNA motifs into the transcriptional unit of a gene or vector. When correctly positioned, the sequences lead to potent inhibition of gene or vector expression, owing to the spontaneous cleavage of the RNA transcript. Administration of either oligonucleotides complementary to regions of the self-cleaving motif or a specific small molecule results in the efficient induction of gene expression, owing to inhibition of self-cleavage of the messenger RNA. Efficient regulation of transgene expression is shown in a variety of mammalian cell lines and live animals. In conjunction with other emerging technologies, this methodology may be particularly applicable to the development of gene regulation systems tailored to any small inducer molecule, and provide a novel means of biological sensing in vivo that may have an important application in the regulated delivery of protein therapeutics.
Yen
Laising
L
Department of Genetics, Harvard Institute of Human Genetics, Harvard Medical School, and Division of Molecular Medicine, Children's Hospital, Boston, Massachusetts 02115, USA.
Svendsen
Jennifer
J
Lee
Jeng-Shin
JS
Gray
John T
JT
Magnier
Maxime
M
Baba
Takashi
T
D'Amato
Robert J
RJ
Mulligan
Richard C
RC
D000241:Adenosine / Q000494:pharmacology; D000818:Animals; D020029:Base Pairing; D001483:Base Sequence; D002384:Catalysis / Q000187:drug effects; D002460:Cell Line; D045744:Cell Line, Tumor; D006224:Cricetinae; D005786:Gene Expression Regulation / Q000187:drug effects; D017930:Genes, Reporter / Q000235:genetics; D005818:Genetic Engineering / Q000379:methods*; D005822:Genetic Vectors / Q000235:genetics; D006801:Humans; D051379:Mice; D008969:Molecular Sequence Data; D009841:Oligonucleotides / Q000737:chemistry / Q000235:genetics / Q000378:metabolism / Q000494:pharmacology; D009928:Organ Specificity; D016337:RNA, Catalytic / Q000037:antagonists & inhibitors* / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D014127:Toyocamycin / Q000494:pharmacology
D009841:Oligonucleotides; D016337:RNA, Catalytic; D012333:RNA, Messenger; D000241:Adenosine; D014127:Toyocamycin