Thermal regulation of beta-galactosidase synthesis using anti-sense RNA directed against the coding portion of the mRNA.
The in vivo production of RNA that is complementary to the mRNA of a particular target gene (anti-sense RNA) appears to be an effective tool for the regulation of genes in Escherichia coli (Coleman, J., Green, P.J., and Inouye, M. (1984) Cell 37, 429-436). These investigators demonstrated that short anti-sense transcripts which are complementary to the ribosome binding site of the target mRNA are overwhelmingly the most effective in blocking protein synthesis. We have constructed plasmids which produce thermally regulated anti-sense transcripts of three regions of the E. coli lac Z gene coding sequence, and have examined the relative effects of these constructs on the synthesis of the lac Z gene product, beta-galactosidase. We conclude that there is a strong correlation between the length of RNA complementarity and the suppression of beta-galactosidase synthesis. Furthermore, a significant inhibition of translation can be obtained when the anti-sense transcript lacks complementarity to the 5' noncoding region of the mRNA, provided that the extent of complementarity with the coding sequence is considerable.
M J
MJ
Ellison
R J
RJ
Kelleher
A
A
Rich
D004262:DNA Restriction Enzymes; D004926:Escherichia coli / Q000201:enzymology* / Q000235:genetics*; D005696:Galactosidases / Q000235:genetics*; D005796:Genes; D005798:Genes, Bacterial; D005809:Genes, Regulator; D007700:Kinetics; D010957:Plasmids; D012329:RNA, Bacterial / Q000235:genetics*; D012333:RNA, Messenger / Q000235:genetics*; D013696:Temperature; D014158:Transcription, Genetic; D001616:beta-Galactosidase / Q000096:biosynthesis / Q000235:genetics*
D012329:RNA, Bacterial; D012333:RNA, Messenger; D004262:DNA Restriction Enzymes; D005696:Galactosidases; D001616:beta-Galactosidase