Technical knockout (TKO) is a method for function-based gene isolation in mammalian cells.
Technical knockout application has resulted in the isolation of several novel genes involved in apoptosis and in the genetic dissection of certain processes within this important biological process.
One concept is the use of nucleic acids as “virtual mutagens” for trapping genes based on their function. This was achieved by transfections with antisense cDNA libraries to randomly inactivate gene expression, followed by a strong phenotypic selection.
The other concept was to develop efficient strategies for introduction and recovery of complex cDNA libraries, to allow multiple rounds of phenotypic selection and rapid screens for identification of functionally relevant genes. To this aim, an expression vector that provides an efficient gene...