RNA aptamers as effective protein antagonists in a multicellular organism.
RNA aptamers selected against proteins can be used to modulate specific protein function. Expression of such reagents in cells and whole organisms could provide a means of dissecting and controlling molecular mechanisms in vivo. We demonstrate that Drosophila B52 protein can be specifically inhibited in vitro and in vivo by a multivalent RNA aptamer. This inhibitory aptamer RNA binds B52 avidly and inhibits B52-stimulated pre-mRNA splicing. It can be expressed in cultured cells and whole animals in a stable form that accumulates up to 10% of total mRNA. It binds B52 in vivo and suppresses all phenotypes caused by B52 overexpression. The strategies presented here should prove general in design and expression of functional and therapeutic RNAs.
Shi
H
H
Department of Molecular Biology and Genetics, Cornell University, Ithaca, NY 14853, USA.
Hoffman
B E
BE
Lis
J T
JT
D000818:Animals; D030801:Animals, Genetically Modified; D001483:Base Sequence; D002478:Cells, Cultured; D003433:Crosses, Genetic; D004330:Drosophila / Q000235:genetics* / Q000378:metabolism; D029721:Drosophila Proteins; D005260:Female; D005838:Genotype; D008297:Male; D008969:Molecular Sequence Data; D009687:Nuclear Proteins / Q000037:antagonists & inhibitors*; D009690:Nucleic Acid Conformation; D009843:Oligoribonucleotides / Q000494:pharmacology*; D010641:Phenotype; D010750:Phosphoproteins / Q000037:antagonists & inhibitors*; D012313:RNA / Q000737:chemistry* / Q000235:genetics*; D012326:RNA Splicing; D000072260:RNA Splicing Factors; D012333:RNA, Messenger / Q000235:genetics*; D016601:RNA-Binding Proteins / Q000037:antagonists & inhibitors; D014158:Transcription, Genetic
D029721:Drosophila Proteins; D009687:Nuclear Proteins; D009843:Oligoribonucleotides; D010750:Phosphoproteins; D000072260:RNA Splicing Factors; D012333:RNA, Messenger; D016601:RNA-Binding Proteins; C070791:B52 protein, Drosophila; D012313:RNA