A Novel Method to Silence Genes in Cryptosporidium

Research output: Chapter in Book/Report/Conference proceedingChapter

1 Scopus citations

Abstract

The parasites of the genus Cryptosporidium are important causes of diarrheal diseases, specifically cryptosporidiosis, worldwide. A major bottleneck for developing drugs and vaccines against cryptosporidiosis is the lack of methods to study gene function in this parasite. Silencing of genes by RNA interference (RNAi) is a powerful method to investigate gene function that has been widely used in the identification of targets for several pathogens. Unfortunately, as Cryptosporidium does not possess the enzymes of the RNAi pathway, its genes cannot be silenced by standard siRNA technology. To circumvent that problem, we have developed a novel strategy to knock down Cryptosporidium genes by reconstituting the effector arm of the siRNA pathway. We have induced silencing of several genes in Cryptosporidium by transfecting parasites with hybrid complexes formed between recombinant human Argonaute (hAgo2) and Cryptosporidium single-stranded RNA (ssRNA). This novel methodology provides an effective strategy to study the role of selected genes in host–parasite interactions, and also can be used to identify potential targets for chemotherapy. The standardized methodology based on this strategy is described in this chapter.

Original languageEnglish (US)
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages193-203
Number of pages11
DOIs
StatePublished - 2020

Publication series

NameMethods in Molecular Biology
Volume2052
ISSN (Print)1064-3745
ISSN (Electronic)1940-6029

Keywords

  • Cryptosporidium
  • Gene silencing
  • RNAi

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

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    Castellanos-Gonzalez, A. (2020). A Novel Method to Silence Genes in Cryptosporidium. In Methods in Molecular Biology (pp. 193-203). (Methods in Molecular Biology; Vol. 2052). Humana Press Inc.. https://doi.org/10.1007/978-1-4939-9748-0_11