title: Silencing of HTLV-1 gag and env genes by small interfering RNAs in HEK 293 cells.
abstract: Since the discovery of RNAi technology, several functional genomic and disease therapy studies have been conducted using this technique in the field of oncology and virology. RNAi-based antiviral therapies are being studied for the treatment of retroviruses such as HIV-1. These studies include the silencing of regulatory, infectivity and structural genes. The HTLV-1 structural genes are responsible for the synthesis of proteins involved in the entry, assembly and release of particles during viral infection. To examine the possibility of silencing HTLV-1 genes gag and env by RNA interference technology, these genes were cloned into reporter plasmids. These vectors expressed the target mRNAs fused to EGFP reporter genes. Three small interference RNAs (siRNAs) corresponding to gag and three corresponding to env were designed to analyze the effect of silencing by RNAi technology. The plasmids and siRNAs were co-transfected into HEK 293 cells. The results demonstrated that the expression of the HTLV-1 gag and env genes decreased significantly in vitro. Thus, siRNAs can be used to inhibit HTLV-1 structural genes in transformed cells, which could provide a tool for clarifying the roles of HTLV-1 structural genes, as well as a therapy for this infection.
authors.affiliation: Universidade de São Paulo, Ribeirão Preto, São Paulo, Brazil.
authors.forename: Rodrigo
authors.initials: R
authors.lastname: Haddad
authors.forename: Simone
authors.initials: S
authors.lastname: Kashima
authors.forename: Evandra Strazza
authors.initials: ES
authors.lastname: Rodrigues
authors.forename: Rochele
authors.initials: R
authors.lastname: Azevedo
authors.forename: Patrícia Vianna Bonini
authors.initials: PV
authors.lastname: Palma
authors.forename: Danielle Aparecida Rosa
authors.initials: DA
authors.lastname: de Magalhães
authors.forename: Marco Antonio
authors.initials: MA
authors.lastname: Zago
authors.forename: Dimas Tadeu
authors.initials: DT
authors.lastname: Covas
mesh_terms: D019677:Artificial Gene Fusion; D015686:Gene Products, env / Q000037:antagonists & inhibitors* / Q000235:genetics; D015683:Gene Products, gag / Q000037:antagonists & inhibitors* / Q000235:genetics; D020868:Gene Silencing; D017930:Genes, Reporter; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D057809:HEK293 Cells; D015368:Human T-lymphotropic virus 1 / Q000235:genetics*; D006801:Humans; D034741:RNA, Small Interfering / Q000235:genetics*
chemical_list: D015686:Gene Products, env; D015683:Gene Products, gag; D034741:RNA, Small Interfering; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins