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A simple and efficient transfection protocol for Cryptosporidium parvum using Polyethylenimine (PEI) and Octaarginine

Published online by Cambridge University Press:  04 May 2020

Tran Nguyen-Ho-Bao ,
Maxi Berberich ,
Wanpeng Zheng ,
Dieter Seebach ,
Arwid Daugschies  and
Faustin Kamena Open the ORCID record for Faustin Kamena [Opens in a new window]
Tran Nguyen-Ho-Bao
Affiliation:
Faculty of Veterinary Medicine, Institute of Parasitology, Centre for Infectious Medicine, University of Leipzig, 04103Leipzig, Germany Department of Veterinary Medicine, College of Agriculture, Can Tho University, 900000Can Tho, Vietnam
Maxi Berberich
Affiliation:
Faculty of Veterinary Medicine, Institute of Parasitology, Centre for Infectious Medicine, University of Leipzig, 04103Leipzig, Germany
Wanpeng Zheng
Affiliation:
Faculty of Veterinary Medicine, Institute of Parasitology, Centre for Infectious Medicine, University of Leipzig, 04103Leipzig, Germany
Dieter Seebach
Affiliation:
Laboratory for Organic Chemistry, Swiss Federal Institute of Technology, ETH-Z, Zurich, 8093 Zürich, Switzerland
Arwid Daugschies
Affiliation:
Faculty of Veterinary Medicine, Institute of Parasitology, Centre for Infectious Medicine, University of Leipzig, 04103Leipzig, Germany
Faustin Kamena*
Affiliation:
Faculty of Veterinary Medicine, Institute of Parasitology, Centre for Infectious Medicine, University of Leipzig, 04103Leipzig, Germany Department of Microbiology and Parasitology, Laboratory for Molecular Parasitology, University of Buea, Cameroon, PO Box 63, Buea, Cameroon
*
Author for correspondence: Faustin Kamena, E-mail: faustin.kamena@uni-leipzig.de
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Abstract

The transfection of Cryptosporidium represents a major challenge, and current protocols are based on electroporation of freshly excysted sporozoites using a rather large amount of plasmid DNA which typically has a very poor yield. In this study, we report a fast and simple protocol for transfection of Cryptosporidium parvum that takes advantage of the DNA condensing power of the poly cationic polymer polyethylenimine (PEI) and the gene delivery property of the short cell-penetrating peptide octaarginine. Our novel protocol requires a very low amount of plasmid DNA and does not necessitate special laboratory equipment to be performed. Transfection appears to be more efficient in oocysts just triggered for excystation than the excysted sporozoites. Altogether, the application of octaarginine with PEI allows efficient transfection. To the best of our knowledge, this is the first report on an electroporation-free protocol for transfection of sporozoites of a Cryptosporidium species.

Keywords

Cryptosporidium parvumoctaargininepolyethyleniminetransfection
Type
Research Article
Information
Parasitology , Volume 147 , Issue 10 , September 2020 , pp. 1065 - 1070
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Copyright
Copyright © The Author(s) 2020. Published by Cambridge University Press

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