Silencing of HTLV-1 gag and env genes by small interfering RNAs in HEK 293 cells |
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Authors: | Haddad Rodrigo Kashima Simone Rodrigues Evandra Strazza Azevedo Rochele Palma Patrícia Vianna Bonini de Magalhães Danielle Aparecida Rosa Zago Marco Antonio Covas Dimas Tadeu |
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Institution: | a Fundação Hemocentro de Ribeirão Preto - Universidade de São Paulo (USP), Rua Tenente Catão Roxo, 2501, Bairro Monte Alegre, Ribeirão Preto, São Paulo, CEP: 14051-140, Brazil b Faculdade de Medicina de Ribeirão Preto - Universidade de São Paulo (USP), Avenida Bandeirantes, 3900, Bairro Monte Alegre, Ribeirão Preto, São Paulo, CEP: 14049-900, Brazil c Faculdade de Ciências Farmacêuticas de Ribeirão Preto - Universidade de São Paulo (USP), Av. do Café, s/n°, Bairro Monte Alegre, Ribeirão Preto, São Paulo, CEP: 14040-903, Brazil |
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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. |
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Keywords: | HTLV-1 Gag Env RNA interference siRNA |
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