Ribozyme and antisense RNAs inhibit coupled transcription translation by binding to rabbit polyribosomes.
The behavior of ribozyme and antisense RNAs was analyzed in a coupled rabbit reticulocyte transcription translation system. Both ribozyme and antisense RNAs were efficiently produced and bound tightly to polyribosomes at 30 degrees C, but did not produce a protein product. Antisense and ribozyme RNA binding depended upon the presence of intact ribosomes, was specific since, plasmid DNA did not associate with either ribosomes or polyribosomes, and was temperature dependent. Ribozyme-specific mRNA cleavage in the coupled system was inferred from translation inhibition studies and was confirmed by primer extension analysis. Thus, ribozyme RNA can inhibit target protein production in the coupled transcription translation system by competing out cellular mRNAs and via targeted message degradation.
Denman
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New York State Institute for Basic Research in Developmental Disabilities, Staten Island, New York 10314, USA.
D000818:Animals; D007700:Kinetics; D010957:Plasmids; D016133:Polymerase Chain Reaction; D011132:Polyribosomes / Q000187:drug effects / Q000378:metabolism*; D014176:Protein Biosynthesis / Q000187:drug effects; D016372:RNA, Antisense / Q000494:pharmacology*; D016337:RNA, Catalytic / Q000378:metabolism*; D011817:Rabbits; D012156:Reticulocytes / Q000378:metabolism; D014158:Transcription, Genetic / Q000187:drug effects
D016372:RNA, Antisense; D016337:RNA, Catalytic