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In order to understand the function of genes, transfection can be used as a method in which artificially prepared knockout and expression constructs are being introduced into cell lines. Since many genes are essential for embryonic development and a homozygous deletion results in non-viable embryos, gene disruption in a cell line by using transfected constructs might be an alternative choice. Electroporation is often used to stably transfect knockout and knockin vectors into DT40 cell line.

The DT40 cell line is derived from ALV (avian leukosis virus)-transformed bursal B cell of chicken. This cell line is suspension cell, and can be cultured in cell culture bottles and in culture plates (6-, 24- and 96-well etc.). Optimal culture temperature for the DT40 is (39 degrees C-) 41 degrees C, because chicken has higher body temperature than human and mouse. Here we summarize protocols for culture medium and condition of the DT40 cell line.