Authors

G A Hovel-Miner¹; H S Kim²; D Schulz²;  ¹ George Washington University, United States;  ² Rockefeller University, United States

Discussion

African trypanosomes are tsetse vector transmitted unicellular parasites that result in devastating human and livestock infections throughout sub-Saharan Africa. In addition, Trypanosoma brucei has demonstrated its usefulness as a model organism toward elucidating fundamental principles in diverse research areas that include cell biology, molecular genetics, and epigenetics. In spite of decades of active discovery in all areas of trypanosome research, more than 60% of the T. brucei genome is annotated as hypothetical genes of unknown function. Further progress in understanding both the pathogenesis and basic biology of Trypanosomes requires the development of versatile approaches for genome-scale functional analysis. Strides have been made using an available RNAi based whole-genome loss-of-function library, yet a complementary gain-of-function library had not been produced. While individual gene studies have proven the usefulness of overexpression studies, whole-genome approaches had been hindered by the complexities of trypanosome gene expression. Using up-to-date T. brucei genome annotations and ribosomal profiling data we have addressed this challenge by generating a PCR amplicon-based overexpression library. The quality of both the plasmid libraries and bloodstream form T. brucei libraries are in the final stages of quality validation. Upon their completion, the libraries will be made broadly available to Trypanosome researchers.