Inhibition of expression of HTLV-1 structural genes mediated by short hairpin RNA in vitro.

BACKGROUND

Human T-lymphotropic virus I (HTLV-1) is associated with the T-cell malignancy known as adult T-cell leukemia/ lymphoma (ATLL) and with a disorder called HTLV-1-associated myelopathy/tropical spastic paraparesis (HAM/TSP). Currently, the treatment of these diseases is based on symptom relief. RNA interference (RNAi) technology has been described as an efficient mechanism for development of new therapeutic methods. Thus, the aim of this study was to evaluate the inhibition of HTLV-1 structural proteins using short hairpin RNAs (shRNAs) expressed by non-viral vectors.

MATERIALS AND

METHODS

Reporter plasmids that express enhanced green fluorescent protein-Gag (EGFP-Gag) and EGFP-Env fusion proteins and vectors that express shRNAs corresponding to the HTLV-1 gag and env genes were constructed. shRNA vectors and reporter plasmids were simultaneously transfected into HEK 293 cells.

RESULTS

Fluorescence microscopy, flow cytometry and real-time PCR showed that shRNAs were effective in inhibiting the fusion proteins.

CONCLUSION

These shRNAs are effective against the expression of structural genes and may provide an approach to the development of new therapeutic agents.