Wannisa KhamaikawinSomphot SaoinSawitree NangolaKoollawat ChupraditSupachai SakkhachornphopSudarat HadpechNattawat OnlamoonAftab A. AnsariSiddappa N. ByrareddyPierre BoulangerSaw See HongBruce E. TorbettChatchai TayapiwatanaChiang Mai UniversityUniversity of PhayaoMahidol UniversityEmory UniversityUniversite de LyonScripps Research Institute2018-11-232018-11-232015-08-25Molecular Therapy - Nucleic Acids. Vol.4, (2015), e249216225312-s2.0-84940053734https://repository.li.mahidol.ac.th/handle/20.500.14594/35392Designed molecular scaffolds have been proposed as alternative therapeutic agents against HIV-1. The ankyrin repeat protein (AnkGAG1D4) and the zinc finger protein (2LTRZFP) have recently been characterized as intracellular antivirals, but these molecules, used individually, do not completely block HIV-1 replication and propagation. The capsid-binder AnkGAG1D4, which inhibits HIV-1 assembly, does not prevent the genome integration of newly incoming viruses. 2LTRZFP, designed to target the 2-LTR-circle junction of HIV-1 cDNA and block HIV-1 integration, would have no antiviral effect on HIV-1-infected cells. However, simultaneous expression of these two molecules should combine the advantage of preventive and curative treatments. To test this hypothesis, the genes encoding the N-myristoylated Myr(+)AnkGAG1D4 protein and the 2LTRZFP were introduced into human T-cells, using a third-generation lentiviral vector. SupT1 cells stably expressing 2LTRZFP alone or with Myr(+)AnkGAG1D4 showed a complete resistance to HIV-1 in viral challenge. Administration of the Myr(+)AnkGAG1D4 vector to HIV-1-preinfected SupT1 cells resulted in a significant antiviral effect. Resistance to viral infection was also observed in primary human CD4+ T-cells stably expressing Myr(+)AnkGAG1D4, and challenged with HIV-1, SIVmac, or SHIV. Our data suggest that our two anti-HIV-1 molecular scaffold prototypes are promising antiviral agents for anti-HIV-1 gene therapy.Mahidol UniversityBiochemistry, Genetics and Molecular BiologyArticleSCOPUS10.1038/mtna.2015.22