| 摘要 |
Methods of identifying inhibitors of retroviral propagation, tRNA used in the methods, and kits, including the tRNA, which can be used in the methods, are disclosed. Methods of treating or preventing retroviral infections by administering an effective amount of the inhibitors, and pharmaceutical compositions including the inhibitors, are also disclosed. The methods involve forming a mixture comprising a linear sequence of a tRNA anticodon stem loop fragment that is not capable of forming a stem-loop, a target nucleic acid molecule capable of binding to the tRNA anticodon stem loop fragment, and a test compound. The mixture is incubated under conditions that allow binding of the tRNA anticodon stem loop fragment and the target nucleic acid molecule in the absence of the test compound. Assays can then be performed that detect whether or not the test compound inhibits the binding of the tRNA anticodon stem loop fragment and the target nucleic acid molecule. |
| 主权项 |
1. A method of identifying an anti-HIV agent, comprising:
forming a mixture comprising a linear sequence of a tRNA anticodon stem loop fragment that is not capable of forming a stem-loop, a target nucleic acid molecule capable of binding to the tRNA anticodon stem loop fragment, and a test compound, wherein the target nucleic acid molecule corresponds to a portion of a retroviral genome involved in: (a) reverse transcription; b) HIV translation of viral RNA to precursor proteins; c) final packaging and assembly; or d) retroviral primer recruitment; incubating the mixture under conditions that allow binding of the tRNA anticodon stem loop fragment and the target nucleic acid molecule in the absence of the test compound; and detecting whether or not the test compound inhibits the binding of the tRNA anticodon stem loop fragment and the target nucleic acid molecule, wherein the absence of binding of the tRNA ASL fragment and the target nucleic acid molecule is indicative of the test compound being: i) an inhibitor of retroviral reverse transcription, ii) an inhibitor of HIV translation of viral RNA to precursor proteins, iii) an inhibitor of HIV final packaging and assembly of new virions by the inhibition of secondary virus spread through the decreasing of viral budding, or iv) an inhibitor of HIV's recruitment of the retroviral primer, human tRNALys3, respectively, wherein the tRNA anticodon stem loop fragment comprises the sequence 5′-GCUXUUAYZCUG (SEQ ID NO: 1), in which the X, Y, and Z refer to modified nucleosides, and optionally comprises a label. |
