Biological detection of specific mRNA molecules by microinjection.
We have used direct microinjection of messenger RNA into individual mouse and human cells to assay for specific translation products. We have been able to detect the synthesis of human fibroblast interferon, thymidine, kinase, hypoxanthine phosphoribosyltransferase, adenine phosphoribosyltransferase, and propionyl-CoA carboxylase in response to injected mRNA. Using the interferon system as a model, we have quantitated interferon synthesis and followed partial purification of interferon mRNA sequences on sucrose density gradients. The methods we have utilized should be applicable to other systems in which sensitive assays exist for gene products and should provide a screening procedure for isolating specific mRNA sequences.
D000228:Adenine Phosphoribosyltransferase / Q000235:genetics; D002262:Carboxy-Lyases / Q000235:genetics; D002460:Cell Line; D006801:Humans; D007041:Hypoxanthine Phosphoribosyltransferase / Q000235:genetics; D007372:Interferons / Q000096:biosynthesis / Q000235:genetics*; D007739:L Cells; D008845:Microinjections; D011422:Propionates; D012333:RNA, Messenger / Q000235:genetics* / Q000378:metabolism; D013937:Thymidine Kinase / Q000235:genetics
D011422:Propionates; D012333:RNA, Messenger; D007372:Interferons; D000228:Adenine Phosphoribosyltransferase; D007041:Hypoxanthine Phosphoribosyltransferase; D013937:Thymidine Kinase; D002262:Carboxy-Lyases