A Novel Method to Silence Genes in Cryptosporidium

    Research output: Chapter in Book/Report/Conference proceedingChapter

    4 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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