Optimization of Polycistronic Anti-CCR5 Artificial microRNA Leads to Improved Accuracy of Its Lentiviral Vector Transfer and More Potent Inhibition of HIV-1 in CD4+ T-Cells
Felix Urusov,
Dina Glazkova,
Denis Omelchenko,
Elena Bogoslovskaya,
Galina Tsyganova,
Katerina Kersting,
German Shipulin,
Vadim Pokrovsky
Affiliations
Felix Urusov
Department of Molecular Diagnostic and Epidemiology, Federal Budget Institution of Science “Central Research Institute of Epidemiology” of The Federal Service on Customers’ Rights Protection and Human Well-being Surveillance, 111123 Moscow, Russia
Dina Glazkova
Department of Molecular Diagnostic and Epidemiology, Federal Budget Institution of Science “Central Research Institute of Epidemiology” of The Federal Service on Customers’ Rights Protection and Human Well-being Surveillance, 111123 Moscow, Russia
Denis Omelchenko
Department of Molecular Diagnostic and Epidemiology, Federal Budget Institution of Science “Central Research Institute of Epidemiology” of The Federal Service on Customers’ Rights Protection and Human Well-being Surveillance, 111123 Moscow, Russia
Elena Bogoslovskaya
Department of Molecular Diagnostic and Epidemiology, Federal Budget Institution of Science “Central Research Institute of Epidemiology” of The Federal Service on Customers’ Rights Protection and Human Well-being Surveillance, 111123 Moscow, Russia
Galina Tsyganova
Department of Molecular Diagnostic and Epidemiology, Federal Budget Institution of Science “Central Research Institute of Epidemiology” of The Federal Service on Customers’ Rights Protection and Human Well-being Surveillance, 111123 Moscow, Russia
Katerina Kersting
Department of Molecular Diagnostic and Epidemiology, Federal Budget Institution of Science “Central Research Institute of Epidemiology” of The Federal Service on Customers’ Rights Protection and Human Well-being Surveillance, 111123 Moscow, Russia
German Shipulin
Department of Molecular Diagnostic and Epidemiology, Federal Budget Institution of Science “Central Research Institute of Epidemiology” of The Federal Service on Customers’ Rights Protection and Human Well-being Surveillance, 111123 Moscow, Russia
Vadim Pokrovsky
Department of Molecular Diagnostic and Epidemiology, Federal Budget Institution of Science “Central Research Institute of Epidemiology” of The Federal Service on Customers’ Rights Protection and Human Well-being Surveillance, 111123 Moscow, Russia
C-C chemokine receptor type 5 (CCR5) is utilized by human immunodeficiency virus (HIV) as a co-receptor for cell entry. Suppression of the CCR5 gene by artificial microRNAs (amiRNAs) could confer cell resistance. In previous work, we created a lentivector that encoded the polycistron of two identical amiRNAs that could effectively suppress CCR5. However, tandem repeats in lentiviral vectors led to deletions of the repeated sequences during reverse transcription of the vector RNA. To solve this problem, we have created a new amiRNA against CCR5, mic1002, which has a different microRNA scaffold and targets a different sequence. Replacing one of the two identical tandem amiRNAs in the polycistron with the mic1002 amiRNA increased the accuracy of its lentiviral vector transfer while retaining its ability to effectively suppress CCR5. A lentiviral vector containing two heterogenic amiRNAs significantly inhibited HIV replication in a vector-transduced human CD4+ lymphocyte culture.
