Constitutive and conditional suppression of exogenous and endogenous genes by anti-sense RNA.
Plasmid DNA directing transcription of the noncoding (anti-sense) DNA strand can specifically inhibit the expression of several test genes as well as normal, endogenous genes. The anti-sense plasmid constructions can be introduced into eukaryotic cells by transfection or microinjection and function in both transient and stable transformation assays. Anti-sense transcripts complementary to as little as 52 bases of 5' untranslated target gene mRNA specifically suppress gene activity as well as, or more efficiently than, anti-sense transcripts directed against the protein coding domain alone. Conditional anti-sense inhibition is accomplished with the use of hormone-inducible promoter sequences. Suppression of endogenous actin gene activity by anti-sense RNA is detected as a decrease in growth rate and as a reduction in the number of actin microfilament cables. These observations suggest that anti-sense RNA may be generally useful for suppressing the expression of specific genes in vivo and may be a potential molecular alternative to classical genetic analysis.
Izant
J G
JG
Weintraub
H
H
D000123:Acetyltransferases / Q000235:genetics; D000199:Actins / Q000235:genetics; D000818:Animals; D002417:Cattle; D002645:Chickens; D015500:Chloramphenicol O-Acetyltransferase; D004274:DNA, Recombinant; D004330:Drosophila; D005796:Genes; D005814:Genes, Viral; D005818:Genetic Engineering; D005822:Genetic Vectors; D005914:Globins / Q000235:genetics; D010957:Plasmids; D012333:RNA, Messenger / Q000235:genetics*; D018139:Simplexvirus / Q000235:genetics; D013489:Suppression, Genetic; D013937:Thymidine Kinase / Q000235:genetics; D014981:Xenopus
D000199:Actins; D004274:DNA, Recombinant; D012333:RNA, Messenger; D005914:Globins; D000123:Acetyltransferases; D015500:Chloramphenicol O-Acetyltransferase; D013937:Thymidine Kinase