Screening methods for the identification of molecular splicing - trans - Pre-efficient

摘要

The present invention, optimum pre - provides methods and compositions for rapid high capacity functional screening to identify splicing molecule (PTM) - trans. Candidate PTM that are designed to mediate splicing reaction - The compositions of the present invention interact target precursor messenger RNA molecule with (target pre-mRNA), transformer resulting in the formation chimeric RNA molecules novel (chimeric RNA) I include PTM expression libraries capable of encoding. Candidate PTM of the invention encodes a reporter molecule in one or more of the other that can be used to select cells that express and coding, and (efficient and specific) optimal PTM part of the reporter molecule in the first can be. The compositions of the present invention, cells expressing the target pre-mRNA encoding the remainder of the reporter molecule to the first well containing. Trans in the presence of (expressed by the library vector) optimal PTM, and cells expressing the target pre-mRNA and (i) PTM expression library screening methods of the present invention, - splicing reaction occurs, whereby at least be brought into contact under conditions that form a chimeric RNA molecule that has been repaired capable of encoding a reporter molecule of one will resu possible to select for cells expressing the reporter molecule is (ii) repair (where expression of the reporter molecule and comprises identifying the best PTM expressed in cells and is (iii) Selection;) indicates that the optimal PTM is present in the selected cells. It is possible to evaluate both the splicing - using more reporter molecules, as well as direct PTM expression, non-specific and specific trans.