Transfection of the African and American Trypanosomes

The protozoan genus Trypanosoma contains several pathogenic parasites of major medical and veterinary importance. These include Trypanosoma brucei subspecies, the causative agents of African trypanosomiasis, and Trypanosoma cruzi , the causative agent of American trypanosomiasis or Chagas’ disease. The development of transformation procedures for these organisms has been a major technical advance. Applications include investigation of the molecular determinants of virulence, functional analysis of potential chemotherapeutic targets, and studies on the mechanisms of gene expression, drug resistance, and developmental regulation. Electroporation is the predominant method used to transfect trypanosomes. The protocols described here are in routine use in our respective laboratories in Amsterdam (T. brucei) and London (T. cruzi) , and have proven to be straightforward and reliable.