Genetic techniques in Trypanosoma cruzi.

It is almost 20 years since genetic manipulation of Trypanosoma cruzi was first reported. In this time, there have been steady improvements in the available vector systems, and the applications of the technology have been extended into new areas. Episomal vectors have been modified to enhance the level of expression of transfected genes and to facilitate the sub-cellular location of their products. Integrative vectors have been adapted to allow the development of inducible expression systems and the construction of vectors which enable genome modification through telomere-associated chromosome fragmentation. The uses of reverse genetic approaches to dissect peroxide metabolism and the mechanisms of drug activity and resistance in T. cruzi are illustrated in this chapter as examples of how the technology has been used to investigate biological function. Although there remains scope to improve the flexibility of these systems, they have made valuable contributions towards exploiting the genome sequence data and providing a greater understanding of parasite biology and the mechanisms of infection.