Design of an Intron-Retained Bioluminescence Reporter and its Application in Imaging of Pre-mRNA Splicing in Living Subjects.

Abstract

Aberrant splicing of precursor messenger RNA (pre-mRNA) can generate abnormal transcripts, and most of the human diseases have been shown to associate with abnormal splicing of pre-mRNA. Conventional methods require sample lysis and thus cannot be used for monitoring pre-mRNA splicing in real time. This chapter guides how to develop an intron-retained bioluminescence (BL) reporter, which simulates the splicing process of pre-mRNA in vitro and in vivo noninvasively. In the following, we illustrate the design and construction of RLuc-intron and the methods of BL experiments in vitro and in vivo. The exemplified results show that our reporter is suitable for high-throughput screening of splicing inhibitors for the therapies of the diseases caused by aberrant splicing.