| 摘要 |
In one aspect, there is described a method for determining the effect of a genetic variation or mutation on the integrity of an RNA transcript comprising the steps of: (a) providing a nucleic acid construct comprising two different reporters separated by an in frame splicing unit, wherein said splicing unit comprises at least two exons separated by at least one intron and wherein at least one exon (eg. the exon downstream of said intron) comprises the genetic variation; (b) transfecting said construct into a cell and/or generating a stable cell line; (c) culturing said cell; and (d) determining the effect of said genetic variation on the integrity of the RNA transcript, wherein a difference in reporter activity in comparison to a cell comprising the nucleic acid construct without the genetic variation is indicative that said genetic variation affects the integrity of the RNA transcript. Assays-such as high throughput assays-are also described for identifying agents (nucleic acids, peptides and small molecules) that modulate the integrity of the RNA transcript and/or are involved in modulating RNA metabolism.
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