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ARS Home » Northeast Area » Beltsville, Maryland (BARC) » Beltsville Agricultural Research Center » Animal Parasitic Diseases Laboratory » Research » Publications at this Location » Publication #385773
Research Project: Foodborne Parasites and their Impact on Food Safety

Location: Animal Parasitic Diseases Laboratory

Title: Comparison of in vitro transformation efficiency methods for Plasmodium falciparum

Author
item WANG, SIQI - Kunming Medical University
item ZENG, WEILIN - Kunming Medical University
item ZHAO, WEI - Kunming Medical University
item XIANG, ZHENG - Kunming Medical University
item ZHAO, HUI - Kunming Medical University
item YANK, QI - Kunming Medical University
item LI, XINXIN - Kunming Medical University
item DUAN, MENGXI - Kunming Medical University
item LI, ZIASONG - Kunming Medical University
item WANG, XUN - Kunming Medical University
item Rosenthal, Benjamin
item YANG, ZHAOQING - Kunming Medical University

Submitted to: Molecular and Biochemical Parasitology
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 11/19/2021
Publication Date: 1/1/2022
Citation: Wang, S., Zeng, W., Zhao, W., Xiang, Z., Zhao, H., Yank, Q., Li, X., Duan, M., Li, Z., Wang, X., Rosenthal, B.M., Yang, Z. 2022. Comparison of in vitro transformation efficiency methods for Plasmodium falciparum. Molecular and Biochemical Parasitology. https://doi.org/10.1016/j.molbiopara.2021.111432.
DOI: https://doi.org/10.1016/j.molbiopara.2021.111432

Interpretive Summary: To better understand and control intracellular parasites, better methods are needed to evaluate gene function. Traditional methods to edit, remove, or introduce genes are hampered by the challenge of getting DNA beyond membranes that protect the host cell and then the parasite. Here, USDA researcher teamed with Chinese investigators to evaluate competing approaches using Plasmodium malaria, a parasite that kills millions of people each year. Among three tested methods, two showed greatest promise. The best method makes use of the parasite's natural system for acquiring and expressing new genetic information. This information will be of interest to biologists, parasitologist, and public health workers seeking solutions to human and veterinary threats.

Technical Abstract: Poor efficiency plagues conventional methods to transfect Plasmodium falciparum with genetic modifications, impeding research aimed at limiting the damage wrought by this agent of severe malaria. Here, we sought and documented improvements, using fluoresce imaging, cell sorting, and drug selection as means to measure efficiency. We achieved transfection efficiencies as high as 10-3. One method (Spontaneous uptake of DNA using the Bio-Rad Gene Pulser Xcell) achieved the highest efficiency (0.48%±0.06%), twice the efficiency of a method using nuclear transfection of ring stages employing the 4D-NucleofectorTM X Kit L. We considered efficiency and the time required to complete drug screening experiments when evaluating the method to genetically modify P. falciparum