Cationic liposome-mediated RNA transfection

Proc Natl Acad Sci U S A. 1989 Aug;86(16):6077-81. doi: 10.1073/pnas.86.16.6077.

Abstract

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.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Animals
  • Cells, Cultured
  • Kinetics
  • Liposomes*
  • Mice
  • Phosphatidylethanolamines*
  • Protein Biosynthesis
  • Quaternary Ammonium Compounds*
  • RNA Caps / genetics*
  • RNA, Messenger / genetics*
  • Templates, Genetic
  • Transcription, Genetic
  • Transfection*

Substances

  • Liposomes
  • Phosphatidylethanolamines
  • Quaternary Ammonium Compounds
  • RNA Caps
  • RNA, Messenger
  • N-(1-(2,3-dioleyloxy)propyl)-N,N,N-trimethylammonium
  • 1,2-dielaidoylphosphatidylethanolamine