Cell based methods and systems for the identification of RNA regulatory sequences and compounds that modulate their functions

摘要

The present invention is directed to methods of screening for a small molecule that modulates the ability of a RNA regulatory sequence to inhibit mRNA translation in a mammalian cell comprising incubating the mammalian cell expressing a reporter mRNA and the RNA regulatory sequence in the presence of the small molecule, wherein the RNA regulatory sequence is not attached to the reporter mRNA; and monitoring a reporter protein signal produced by the reporter mRNA, wherein a change in the signal of the reporter protein in the presence of the small molecule compared to the reporter protein signal in the absence of the small molecule indicates the small molecule modulates the ability of the RNA regulatory sequence to inhibit mRNA translation.

主权项

1. A method of identifying a small molecule compound that inhibits the ability of an hepatitis C virus-internal ribosome entry site (HCV-IRES) mRNA regulatory sequence transcript fragment encoded by a sequence consisting of SEQ ID NO: 26 from inhibiting mRNA translation in a mammalian cell culture comprising
incubating the mammalian cell expressing a reporter mRNA transcript and the mRNA regulatory sequence transcript fragment in the presence of said small molecule compound, wherein said mRNA regulatory sequence transcript fragment is not attached to said reporter mRNA; and wherein said HCV-IRES mRNA regulatory sequence transcript fragment inhibits translation of the reporter mRNA transcript in said mammalian cell in the absence of said small molecule compound; and monitoring a reporter protein signal produced by said reporter mRNA transcript in the presence of said small molecule compound, wherein an increase in the signal of the reporter protein in the presence of said small molecule compound compared to the reporter protein signal in the absence of said small molecule compound indicates said small molecule compound inhibits the ability of said mRNA regulatory sequence transcript fragment to inhibit translation of the reporter mRNA transcript.