Authors
Khadijeh Hashemi and Roland Brock
Department
Dept. of Medical BioSciences, Radboud university medical center, Geert Grooteplein 28, 6525 GA Nijmegen, The Netherlands,
Introduction
The analysis of protein function through transient expression is commonplace in cell- and
molecular biology. A general concern are non-physiological effects through overexpression
which may initiate compensatory or non-endogenous pathways. The chance for
compensatory pathways will increase with the time elapsing between transfection and
observation. Moreover, highly heterogenous expression levels across the cell population may
compromise biochemical analyses, when for example protein interactions are probed by co-immunoprecipitation. Thus, the most reliable results are to be expected if all cells show
uniform expression levels within physiological ranges. Here, we compared the kinetics and
uniformity of expression for cells transfected either with eGFP mRNA or a plasmid coding for
eGFP. For expression, plasmids need to enter the cell nucleus which can only occur for dividing
cells. By comparison, mRNA can also be expressed in non-dividing cells.
