Chapter title |
Expression of HTLV-1 Genes in T-Cells Using RNA Electroporation
|
---|---|
Chapter number | 12 |
Book title |
Human T-Lymphotropic Viruses
|
Published in |
Methods in molecular biology, January 2017
|
DOI | 10.1007/978-1-4939-6872-5_12 |
Pubmed ID | |
Book ISBNs |
978-1-4939-6870-1, 978-1-4939-6872-5
|
Authors |
Mariangela Manicone, Francesca Rende, Ilaria Cavallari, Andrea K. Thoma-Kress, Vincenzo Ciminale, Manicone, Mariangela, Rende, Francesca, Cavallari, Ilaria, Thoma-Kress, Andrea K., Ciminale, Vincenzo |
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. |
Mendeley readers
Geographical breakdown
Country | Count | As % |
---|---|---|
Unknown | 5 | 100% |
Demographic breakdown
Readers by professional status | Count | As % |
---|---|---|
Student > Master | 2 | 40% |
Researcher | 1 | 20% |
Unknown | 2 | 40% |
Readers by discipline | Count | As % |
---|---|---|
Medicine and Dentistry | 3 | 60% |
Agricultural and Biological Sciences | 1 | 20% |
Unknown | 1 | 20% |