Abstract
RNAi experiments are ubiquitously used in cell biology and are achieved by transfection of small interfering RNAs (siRNAs) into cells using a transfection reagent. These results in knock‐down of proteins of interest, and the phenotypic consequences are then analyzed. It is reported here that two common RNA interference (RNAi) transfection reagents, DharmaFECT 1 and INTERFERin, in mock transfections using non‐targeting siRNAs, cause alterations in the lipidome of HeLa cells. Some lipids change in response to both, presumably chemically different, transfection reagents, while other lipid species change only in response to one of the reagents. While the functional implications of these lipidomic alterations remain to be investigated, the authors' experiments suggest that it is important to use appropriate mock transfection controls during RNAi experiments, ideally complemented by an orthogonal perturbation, especially when investigating membrane‐associated phenomena.
Highlights
RNA interference (RNAi) experiments are ubiquitously used in cell biology and are achieved by Membrane-associated proteins play many important roles in cells, and it transfection of small interfering RNAs into cells using a transfection is becoming increasingly clear that the reagent
Some lipids change in response to understood, it is clear that cells can reboth, presumably chemically different, transfection reagents, while other lipid spond to and adapt their lipidomes when species change only in response to one of the reagents
While the functional implications of these lipidomic alterations remain to be investigated, the authors’ experiments suggest that it is important to use appropriate mock transfection controls during RNAi experiments, ideally complemented by an exposed to different stimuli and many studies have explored lipidome changes in response to protein perturbation by RNAi
Summary
While lipid biosynthesis is complex and poorly alterations in the lipidome of HeLa cells. The process of siRNA transfection, in the absence of any protein depletion, affected the lipid composition of cells. RNA interference (RNAi) is very commonly used in cell biology on the experimental design of studies involving membrane proto deplete cellular proteins by destroying mRNA encoding a teins and lipids.
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