Importance of Liposome Complexing Volume in Transfection Optimization

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...

preparation and in vitro transfection of modified cationic liposome-linear polyethyleneimine-dna nanocomplexes for gene delivery

Optimization of SW480 Colon Cancer Cells Transfection with Lipofectamine 2000

ABSTRACT           Background and Objectives: Nonviral carriers including those based on synthetic cationic lipids, offer several advantages over the viral counterparts. These carriers are able to form complexes with nucleic acids and deliver genes into the cells via the cellular endocytosis pathway, without significant toxicity. The level of transg...

Liposome-mediated RNA transfection should be used with caution.

Liposome-mediated RNA transfection appears to present a number of advantages for studying the metabolism of reporter mRNAs in mammalian cells. This method is also widely used to transfect siRNAs. Here we describe results indicating that reporter mRNAs introduced into HeLa cells by liposomes do not present the expected behaviors. Namely, the stability of reporter mRNAs was independent of the pre...

Automated Optimization of Cell Transfection

Cell transfection is an essential technique for interrogating cellular pathways. Determining the optimal conditions for introducing nucleic acids into cells can be a time consuming endeavor at the initiation of a cell biology experiment or screen. Here we demonstrate how the Biomek FXP Workstation can be used to identify the optimal conditions for siRNA transfection in a single experiment. Util...