Generating and maintaining transgenic Cryptosporidium parvum parasites

Mattie C. Pawlowic, Sumiti Vinayak, Adam Sateriale, Carrie F. Brooks, Boris Striepen

Research output: Contribution to journalArticle

11 Citations (Scopus)

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.

Original languageEnglish
Pages (from-to)20B.2.1-20B.2.32
JournalCurrent Protocols in Microbiology
Volume46
Issue number1
DOIs
Publication statusPublished - 15 Aug 2017

Keywords

  • (3-5) CRISPR
  • Cryptosporidium
  • Diarrheal disease
  • Genetics
  • Transfection

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