HIV 201 HTS Assay
The Trana HIV 201 High-Throughput (HTS) Assay – designed to identify compounds that inhibit the use of tRNA by HIV – has the ability to select compounds with anti-HIV bioactivity. Because tRNA is essential for HIV replication, disruption of the virus’ ability to use tRNA would represent a novel target for anti-HIV drug therapy.
Downloads
Trana HIV 201 Assay Backgrounder(PDF)
The Role of tRNA in HIV Replication (PDF)
Trana HIV 201 Assay Technical FAQs (PDF)
Scientists at Trana Discovery have developed the patented technology that forms the basis for the assay used to screen molecular libraries for tRNA inhibitors. The technology centers on the anticodon stem loop (ASL) of tRNA and the importance of nucleotide modifications within the ASL. The ability to synthesize copies or mimics of the ASL with the modifications, just as they occur in nature, is what overcomes previous barriers to high-throughput screening and enables further research and the application of Trana Discovery technology to methodically search for compounds that inhibit interactions with tRNA.
The Trana HIV 201 assay targets interactions in specific steps of the life cycle of the HIV virus – that target is the complex formed in the pre-binding complex of reverse transcription. This occurs when the HIV genomic RNA binds with a hijacked, restructured form of the Human tRNA Lys. The restructured Human tRNALys3SUU forms a complex with the HIV genome RNA during HIV replication. HIV proteins “unwind” tRNA from its form used by Humans, depicted above; the restructured form will stabilize binding to viral genomic RNA. The interactions at the Anticodon Stem-Loop Binding Sites are dependent on modified bases of the sequence in the Human tRNA Lys.
At the High Throughput Screening Center at Southern Research, using the HIV 201 assay, several compounds were identified from Southern Research’s large molecular library that inhibited the assay. In a cell-based study that followed, a subset of these compounds was examined for their inhibition qualities in the replication of the HIV-1 Ba-L virus using live Peripheral Blood Mononuclear Cells (PBMC) and found to have anti-HIV activity. Further testing revealed that antiviral activity was not due to inhibition of reverse transcriptase, the mechanism of action of many of the currently available HIV therapies. Although not definitively confirmed, these early findings suggest that the Trana HIV 201 assay has the ability to select anti-HIV bioactive compounds with a novel mechanism of action.
View the Latest Test Results
Trana Discovery HIV 201 Assay Update from Trana Discovery on Vimeo.
Overview of the Trana HIV 201 High-Throughput (HTS) Assay
Trana Discovery HIV 201 Assay Video from Trana Discovery on Vimeo.
Trana Discovery HIV Video | Produced by LemonStripe | Work for this project was performed under the DAIDS, NIAID contract N01-AI-70042; Roger Miller, Project Officer.
