Expression of HTLV-1 Genes in T-Cells Using RNA Electroporation

Manicone M, Rende F, Cavallari I, Thoma-Kreß A, Ciminale V (2017)

Publication Type: Journal article

Publication year: 2017

Journal

Methods in Molecular Biology Humana Press

Book Volume: 1582

Pages Range: 155-170

DOI: 10.1007/978-1-4939-6872-5_12

Abstract

Human T-cell leukemia virus type 1 (HTLV-1) infects about 20 million people world-wide. Around 5% of the infected individuals develop adult T-cell leukemia (ATL) or a neurological disease termed tropical spastic paraparesis (TSP) after a clinical latency of years to decades. Through the use of two promoters and alternative splicing HTLV-1 expresses at least 12 different proteins. HTLV-1 establishes a life-long persistent infection by inducing the clonal expansion of infected cells, a property largely ascribed to the viral genes Tax and HBZ. However, the fact that ATL arises in a minority of infected individuals after a long clinical latency suggests the existence of factors counterbalancing the oncogenic potential of HTLV-1 in the context of natural infection.To study the role of the different HTLV-1 gene products in the HTLV-1 life cycle, we optimized a transfection protocol for primary T-cells using an approach based on the electroporation of in vitro-transcribed RNA. Results showed that the RNA transfection technique combines a high transfection efficiency with low toxicity, not only in Jurkat T-cells but also in primary T-cells. These findings suggest that RNA electroporation is preferable for experiments aimed at investigating the role of HTLV-1 gene products in the context of primary T-cells, which represent the main target of HTLV-1 in vivo.

Authors with CRIS profile

Andrea Thoma-Kreß Institute of Clinical and Molecular Virology

Involved external institutions

University of Padua / Universita degli Studi di Padova IT Italy (IT) Istituto di ricovero e cura a carattere scientifico (IRCCS) IT Italy (IT)

How to cite

APA:

Manicone, M., Rende, F., Cavallari, I., Thoma-Kreß, A., & Ciminale, V. (2017). Expression of HTLV-1 Genes in T-Cells Using RNA Electroporation. Methods in Molecular Biology, 1582, 155-170. https://dx.doi.org/10.1007/978-1-4939-6872-5_12

MLA:

Manicone, Mariangela, et al. "Expression of HTLV-1 Genes in T-Cells Using RNA Electroporation." Methods in Molecular Biology 1582 (2017): 155-170.

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