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A Trypanosoma brucei ORFeome-based Gain-of-Function Library reveals novel genes associated with melarsoprol resistance

M Carter, HS Kim, S Gomez, S Gritz, S Larson, D Schulz, View ORCID ProfileGA Hovel-Miner
doi: https://doi.org/10.1101/849042
M Carter
1The George Washington University, Department of Microbiology, Immunology, and Tropical Medicine, 2300 Eye St., Washington, D.C, 20037
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HS Kim
2The Public Health Research Institute at the International Center for Public Health New Jersey Medical School – Rutgers, The State University of New Jersey, 225 Warren Street, Newark, NJ 07103-3535,
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  • For correspondence: heesook@rutgers.edu
S Gomez
1The George Washington University, Department of Microbiology, Immunology, and Tropical Medicine, 2300 Eye St., Washington, D.C, 20037
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S Gritz
1The George Washington University, Department of Microbiology, Immunology, and Tropical Medicine, 2300 Eye St., Washington, D.C, 20037
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S Larson
1The George Washington University, Department of Microbiology, Immunology, and Tropical Medicine, 2300 Eye St., Washington, D.C, 20037
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D Schulz
3Harvey Mudd College, Department of Biology, 1250 N. Dartmouth Ave., Claremont, CA 91711,
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  • For correspondence: ghovel_miner@gwu.edu dschulz@hmc.edu
GA Hovel-Miner
1The George Washington University, Department of Microbiology, Immunology, and Tropical Medicine, 2300 Eye St., Washington, D.C, 20037
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  • ORCID record for GA Hovel-Miner
  • For correspondence: ghovel_miner@gwu.edu
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ABSTRACT

Trypanosoma brucei is an early branching protozoan that causes Human and Animal African Trypanosomiasis. Forward genetics approaches are powerful tools for uncovering novel aspects of Trypanosomatid biology, pathogenesis, and therapeutic approaches against trypanosomiasis. Here we have generated a T. brucei ORFeome consisting of over 90% of the targeted genome and used it to make an inducible Gain-of-Function library for broadly applicable forward genetic screening. Using a critical drug of last resort, melarsoprol, we conducted a proof of principle genetic screen. Hits arising from this screen support the significance of trypanothione, a key player in redox metabolism, as a target of melarsoprol and implicate novel proteins of the flagellum and mitochondria in drug resistance. This study has produced two powerful new genetic tools for kinetoplastida research, which are expected to promote major advances in kinetoplastida biology and therapeutic development in the years to come.

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Posted November 20, 2019.
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A Trypanosoma brucei ORFeome-based Gain-of-Function Library reveals novel genes associated with melarsoprol resistance
M Carter, HS Kim, S Gomez, S Gritz, S Larson, D Schulz, GA Hovel-Miner
bioRxiv 849042; doi: https://doi.org/10.1101/849042
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A Trypanosoma brucei ORFeome-based Gain-of-Function Library reveals novel genes associated with melarsoprol resistance
M Carter, HS Kim, S Gomez, S Gritz, S Larson, D Schulz, GA Hovel-Miner
bioRxiv 849042; doi: https://doi.org/10.1101/849042

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