Silencing of HTLV-1 gag and env genes by small interfering RNAs in HEK 293 cells

被引:2
作者
Haddad, Rodrigo [1 ,2 ]
Kashima, Simone [1 ,2 ,3 ]
Rodrigues, Evandra Strazza [1 ]
Azevedo, Rochele [1 ]
Bonini Palma, Patricia Vianna [1 ]
Rosa de Magalhaes, Danielle Aparecida [1 ]
Zago, Marco Antonio [1 ,2 ]
Covas, Dimas Tadeu [1 ,2 ]
机构
[1] Univ Sao Paulo, Fundacao Hemoctr Ribeirao Preto, BR-14051140 Sao Paulo, Brazil
[2] Univ Sao Paulo, Fac Med Ribeirao Preto, BR-14049900 Sao Paulo, Brazil
[3] Univ Sao Paulo, Fac Ciencias Farmaceut Ribeirao Preto, BR-14040903 Sao Paulo, Brazil
基金
巴西圣保罗研究基金会;
关键词
HTLV-1; Gag; Env; RNA interference; siRNA; ANTIVIRAL DEFENSE-MECHANISM; HEPATITIS-B-VIRUS; LEUKEMIA-VIRUS; T-CELLS; EXPRESSION; INHIBITION; REPLICATION; TAX; INFECTION;
D O I
10.1016/j.jviromet.2011.01.012
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
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. (C) 2011 Elsevier B.V. All rights reserved.
引用
收藏
页码:92 / 98
页数:7
相关论文
共 47 条
  • [1] Phenotypic silencing of cytoplasmic genes using sequence-specific double-stranded short interfering RNA and its application in the reverse genetics of wild type negative-strand RNA viruses
    Bitko V.
    Barik S.
    [J]. BMC Microbiology, 1 (1) : 1 - 11
  • [2] Nedd4.1-mediated ubiquitination and subsequent recruitment of Tsg101 ensure HTLV-1 Gag trafficking towards the multivesicular body pathway prior to virus budding
    Blot, V
    Perugi, F
    Gay, B
    Prévost, MC
    Briant, L
    Tangy, F
    Abriel, H
    Staub, O
    Dokhélar, MC
    Pique, C
    [J]. JOURNAL OF CELL SCIENCE, 2004, 117 (11) : 2357 - 2367
  • [3] Specific inhibition of gene expression by small double-stranded RNAs in invertebrate and vertebrate systems
    Caplen, NJ
    Parrish, S
    Imani, F
    Fire, A
    Morgan, RA
    [J]. PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2001, 98 (17) : 9742 - 9747
  • [4] Cooper Sarah A, 2009, Pract Neurol, V9, P16, DOI 10.1136/jnnp.2008.167155
  • [5] Duplexes of 21-nucleotide RNAs mediate RNA interference in cultured mammalian cells
    Elbashir, SM
    Harborth, J
    Lendeckel, W
    Yalcin, A
    Weber, K
    Tuschl, T
    [J]. NATURE, 2001, 411 (6836) : 494 - 498
  • [6] Potent and specific genetic interference by double-stranded RNA in Caenorhabditis elegans
    Fire, A
    Xu, SQ
    Montgomery, MK
    Kostas, SA
    Driver, SE
    Mello, CC
    [J]. NATURE, 1998, 391 (6669) : 806 - 811
  • [7] RNA interference against viruses: strike and counterstrike
    Haasnoot, Joost
    Westerhout, Ellen M.
    Berkhout, Ben
    [J]. NATURE BIOTECHNOLOGY, 2007, 25 (12) : 1435 - 1443
  • [8] Role of human T-cell leukemia virus type I Tax in expression of the human telomerase reverse transcriptase (hTERT) gene in human T-cells
    Hara, Toshifumi
    Matsumura-Arioka, Yuuki
    Ohtani, Kiyoshi
    Nakamura, Masataka
    [J]. CANCER SCIENCE, 2008, 99 (06) : 1155 - 1163
  • [9] Hayafune Masaaki, 2006, Antiviral Chemistry & Chemotherapy, V17, P241
  • [10] Tax-inducible production of CC chemokine ligand 22 by human T cell leukemia virus type 1 (HTLV-1)-Infected T cells promotes preferential transmission of HTLV-1 to CCR4-expressing CD4+ T cells
    Hieshima, Kunio
    Nagakubo, Daisuke
    Nakayama, Takashi
    Shirakawa, Aiko-Konno
    Jin, Zhe
    Yoshie, Osamu
    [J]. JOURNAL OF IMMUNOLOGY, 2008, 180 (02) : 931 - 939