Cell-Based Microarrays: Recent Advances for Gene Function Analyses

Reverse transfected cell microarrays (RTCM) are a powerful tool for the systematic analyses of gene functions. With this technology more than a thousand different nucleic acids can be transfected into eukaryotic cells in parallel on a single glass slide. This allows high-throughput analyses of gene functions using gain-of-function, loss-of-function, and mutation approaches. RTCM paved the way for genome-wide gene function analyses in order to determine gene functions involved in the molecular regulation of cell phenotypes in physiologic and pathophysiologic processes. Since RTCM was first introduced in 2001, the technique has been well established. Themethod was successfully used in several genome-wide and large-scale screenings, and novel analysis methods to detect gene functions have been developed. This chapter will summarize the most recent technological developments in the usage of RTCM, including optimization of (1) transfection efficiency, (2) reporter systems and automated data acquisition, (3) spotting density with decreased cross-contamination, and (4) the new development of assays to screen for paracrine gene effects.