Transfection is a method for delivering nucleic acids, either DNA or RNA, into animal cells in vitro. In applications such as protein production, the main purpose is to produce protein; consequently any effects on endogenous gene expression are generally irrelevant, unless the changes relate directly to loss of the cell's ability to produce protein, i.e., cell death. In contrast, in applications where the biochemical pathways are of interest, changes within the cell can alter the interpretation of the data. Unintended up- or down-regulation of genes within transfected cells is termed an off-target effect. This may be caused by the reagent itself, the vector, or the process of transfection. Such effects, if not carefully evaluated, may lead to misinterpretation of the transfection results and to misleading conclusions. Critically important in transfection experiments is the selection of the appropriate control. more