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Generating and Maintaining Transgenic Cryptosporidium parvum Parasites
Mattie C Pawlowic1, Sumiti Vinayak1, Adam Sateriale1
1Center for Tropical and Emerging Global Diseases, University of Georgia, Athens, Georgia.
Abstract:
The apicomplexan parasite Cryptosporidium is a leading cause of diarrheal disease and an important contributor to overall global child mortality. We currently lack effective treatment and immune prophylaxis. Recent advances now permit genetic modification of this important pathogen. We expect this to produce rapid advances in fundamental as well as translational research on cryptosporidiosis. Here we outline genetic engineering for Cryptosporidium in sufficient detail to establish transfection in any laboratory that requires access to this key technology. This chapter details the conceptual design consideration, as well as the experimental steps required to transfect, select, and isolate transgenic parasites. We also provide detail on key in vitro and in vivo assays to detect, validate, and quantify genetically modified Cryptosporidium parasites. © 2017 by John Wiley & Sons, Inc.
Insights
Genetic engineering of Cryptosporidium, a parasite causing diarrheal disease, is now feasible. This breakthrough enables new research into treatments and prevention for cryptosporidiosis, improving child health outcomes.
Area of Science:
- Parasitology
- Molecular Biology
- Infectious Diseases
Background:
- Cryptosporidium is a major cause of diarrheal illness and child mortality globally.
- Current treatments and preventive strategies for cryptosporidiosis are limited.
- Genetic modification of Cryptosporidium was previously challenging.
Purpose of the Study:
- To provide a detailed protocol for genetic engineering of Cryptosporidium.
- To enable researchers to establish transfection technology in their laboratories.
- To facilitate fundamental and translational research on cryptosporidiosis.
Main Methods:
- Detailed conceptual design considerations for genetic engineering.
- Experimental steps for transfection, selection, and isolation of transgenic parasites.
- In vitro and in vivo assays for detection and validation of genetically modified parasites.
Main Results:
- Establishment of a reliable method for genetic engineering of Cryptosporidium.
- Demonstration of successful transfection, selection, and isolation of modified parasites.
- Validation of assays for quantifying genetically modified Cryptosporidium.
Conclusions:
- Genetic engineering of Cryptosporidium is now an accessible technology.
- This advancement is expected to accelerate research into cryptosporidiosis treatments and prevention.
- The provided methods will empower broader research efforts to combat this pathogen.
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