title: Preassembled Single-Stranded RNA-Argonaute Complexes: A Novel Method to Silence Genes in Cryptosporidium.
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.
authors.affiliation: Infectious Disease Division, Department of Internal Medicine, University of Texas Medical Branch, Galveston.
authors.forename: Alejandro
authors.initials: A
authors.lastname: Castellanos-Gonzalez
authors.affiliation: Infectious Disease Division, Department of Internal Medicine, University of Texas Medical Branch, Galveston.
authors.forename: Nicolas
authors.initials: N
authors.lastname: Perry
authors.affiliation: Infectious Disease Division, Department of Internal Medicine, University of Texas Medical Branch, Galveston.
authors.forename: Samantha
authors.initials: S
authors.lastname: Nava
authors.affiliation: Infectious Disease Division, Department of Internal Medicine, University of Texas Medical Branch, Galveston.
authors.forename: A Clinton
authors.initials: AC
authors.lastname: White
mesh_terms: D060565:Argonaute Proteins / Q000235:genetics* / Q000378:metabolism; D003457:Cryptosporidiosis / Q000469:parasitology; D003458:Cryptosporidium / Q000235:genetics* / Q000378:metabolism; D020868:Gene Silencing; D006790:Host-Parasite Interactions / Q000235:genetics*; D006801:Humans; D016053:RNA, Protozoan / Q000235:genetics* / Q000378:metabolism; D034741:RNA, Small Interfering / Q000235:genetics* / Q000378:metabolism; D014162:Transfection
chemical_list: C466422:AGO2 protein, human; D060565:Argonaute Proteins; D016053:RNA, Protozoan; D034741:RNA, Small Interfering
keywords: Argonaute; Cryptosporidium; Cryptosporidium parvum; cryptosporidiosis; siRNA; silencing