Preassembled Single-Stranded RNA-Argonaute Complexes: A Novel Method to Silence Genes in Cryptosporidium

Alejandro Castellanos, Nicolas Perry, Samantha Nava, A. Clinton White

Research output: Contribution to journalArticle

11 Scopus citations

Abstract

Cryptosporidiosis is a common cause of diarrhea morbidity and mortality worldwide. Research progress on this infection has been slowed by lack of methods to genetically manipulate Cryptosporidium parasites. Small interfering RNA (siRNA) is widely used to study gene function, but Cryptosporidium species lack the enzymes necessary to process siRNA. By preassembling complexes with the human enzyme Argonaute 2 (hAgo2) and Cryptosporidium single-stranded RNA (ssRNA), we induced specific slicing in Cryptosporidium RNA targets. We demonstrated the reduction in expression of target genes at the mRNA and protein levels by transfecting live parasites with ssRNA-hAgo2 complexes. Furthermore we used this method to confirm the role of selected molecules during host cell invasion. This novel method provides a novel means of silencing Cryptosporidium genes to study their role in host-parasite interactions and as potential targets for chemotherapy.

Original languageEnglish (US)
Pages (from-to)1307-1314
Number of pages8
JournalJournal of Infectious Diseases
Volume213
Issue number8
DOIs
StatePublished - Apr 15 2016

Keywords

  • Argonaute
  • cryptosporidiosis
  • Cryptosporidium
  • Cryptosporidium parvum
  • silencing
  • siRNA

ASJC Scopus subject areas

  • Infectious Diseases
  • Immunology and Allergy

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