Cationic liposome-mediated RNA transfection.
We have developed an efficient and reproducible method for RNA transfection, using a synthetic cationic lipid, N-[1-(2,3-dioleyloxy)propyl]-N,N,N-trimethylammonium chloride (DOTMA), incorporated into a liposome (lipofectin). Transfection of 10 ng to 5 micrograms of Photinus pyralis luciferase mRNA synthesized in vitro into NIH 3T3 mouse cells yields a linear response of luciferase activity. The procedure can be used to efficiently transfect RNA into human, rat, mouse, Xenopus, and Drosophila cells. Using the RNA/lipofectin transfection procedure, we have analyzed the role of capping and beta-globin 5' and 3' untranslated sequences on the translation efficiency of luciferase RNA synthesized in vitro. Following transfection of NIH 3T3 cells, capped mRNAs with beta-globin untranslated sequences produced at least 1000-fold more luciferase protein than mRNAs lacking these elements.
Malone
R W
RW
Molecular Biology and Virology Laboratory, Salk Institute, San Diego, CA 92138.
Felgner
P L
PL
Verma
I M
IM
D000818:Animals; D002478:Cells, Cultured; D007700:Kinetics; D008081:Liposomes; D051379:Mice; D010714:Phosphatidylethanolamines; D014176:Protein Biosynthesis; D000644:Quaternary Ammonium Compounds; D012315:RNA Caps / Q000235:genetics*; D012333:RNA, Messenger / Q000235:genetics*; D013698:Templates, Genetic; D014158:Transcription, Genetic; D014162:Transfection
D008081:Liposomes; D010714:Phosphatidylethanolamines; D000644:Quaternary Ammonium Compounds; D012315:RNA Caps; D012333:RNA, Messenger; C054005:N-(1-(2,3-dioleyloxy)propyl)-N,N,N-trimethylammonium; C020888:1,2-dielaidoylphosphatidylethanolamine