Cationic liposome-mediated RNA transfection.

TLDR

The study uses the RNA/lipofectin transfection procedure to examine how capping and beta‑globin untranslated sequences affect luciferase RNA translation in vitro. The procedure efficiently delivers RNA to human, rat, mouse, Xenopus, and Drosophila cells, enabling such translational analyses. Using DOTMA‑lipofectin, transfection of 10 ng–5 µg luciferase mRNA into NIH 3T3 cells produced a linear activity response, and capped mRNAs with beta‑globin untranslated regions yielded over 1000‑fold higher protein production.

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.