Antisense and virus trans-activator decoy approaches of inhibiting replication of human T-cell lymphotropic virus type 1 (HTLV-1)

Contents: Introduction 8 The aim and the objectives of the present study 9 Literature review 11 Biology o/HTLV-1 11 Structure of the virion 11 HTLV-1 life cycle 12 Genome structure and encoded proteins 14 The viral oncoprotein - Tax 19 Target cell and receptor 22 HTLV-1 associated diseases 23 Therapeutic nucleic acids and the mechanisms of their action 29 Antisense nucleic acids 29 RNA interference 30 Ribozymes 31 Transcription factor decoy 31 Construction of therapeutic nucleic acids' expression vectors 32 Choice of a promoter 32 Choice of the target 33 Studies in vitro 34 Experiments in vivo 36 Conclusions and future prospects 37 Materials and methods 38 Results 46 Establishment of experimental model of HTLV-1 infection in monolayer cell line HOS 46 Construction of the plasmids carrying asRNA genes targeted at HTLV-1 LTR U3 and pX region and Tax decoy HTLV-1 LTR U3 sequence 60 Investigation of the effect of HTLV-1 asRNA and Tax decoy plasmids on HTLV-1 replication in RaHOS cells 70 Investigation of the effect of HTLV-1 LTR U3 asRNA and Tax decoy plasmids with inserted marker neo gene on virus replication in HTLV-1 transformed rabbit lymphoid cell line Ra-1 and human T-cell line MT-2 82 Discussion 91 Conclusions 97 Acnowledgements 98 References 9