Cell-Based Microarrays of Infectious Adenovirus Encoding Short Hairpin RNA (shRNA)

Genome-wide screening procedures have developed into a useful tool for assigning new functions to known proteins or for identifying new interplayers in cell metabolism, especially under pathological conditions. Since primary cells reflect the physiological situation more closely than transformed cell lines, their employment in such screenings is highly desirable. A difficulty to overcome – besides the shortage in cell supply – is that primary cells are less amenable to classical methods of genetic manipulation such as lipofection or electroporation. By using adenovirus as the vehicle for genetic manipulation, efficient transfer of genetic information can be achieved without drastic effects on cell viability, and by using a microarray format even small amounts of cells suffice to perform a screen.