We developed a technique for mutagenesis of Dictyostelium based on the observation that antisense works well in this system to repress expression of selected gene products.

The cDNA library can be made from any selected developmental stage of cell-type, and the promoter can also be developmental-stage or cell-type specific.

For instance, for one project we were interested in finding genes expressed in vegetative growth that affect cell-type choice during late development, so we used a vegetative promoter and a vegetative cDNA library.

After identifying an interesting mutation, PCR is used to immediately obtain the antisense cDNA, which is then sequenced.