Imaging of pre-mRNA splicing in living subjects using a genetically encoded luciferase reporter

Yaru Shi; Wei Liu; Haifeng Zheng; Zhiqiang Li; Xiaorui Shi; Shixuan Cai; Zhiqiang Jiao; Wenjie Mao; Jinrong Xie; Jie Tian; Fu Wang

doi:10.1364/BOE.9.000518

Imaging of pre-mRNA splicing in living subjects using a genetically encoded luciferase reporter

Biomed Opt Express. 2018 Jan 8;9(2):518-528. doi: 10.1364/BOE.9.000518. eCollection 2018 Feb 1.

Authors

Yaru Shi^{1

2}, Wei Liu^{3

2}, Haifeng Zheng¹, Zhiqiang Li¹, Xiaorui Shi¹, Shixuan Cai¹, Zhiqiang Jiao¹, Wenjie Mao¹, Jinrong Xie¹, Jie Tian^{1

4

5}, Fu Wang¹

Affiliations

¹ Engineering Research Center of Molecular and Neuro Imaging, Ministry of Education, School of Life Science and Technology, Xidian University, Xi'an, Shaanxi 710071, China.
² Equal contribution.
³ Department of Hepatobiliary Surgery, Xijing Hospital, The Fourth Military Medical University, Xi'an, Shaanxi 710032, China.
⁴ CAS Key Laboratory of Molecular Imaging, Institute of Automation, Chinese Academy of Sciences, Beijing 100190, China.
⁵ jie.tian@ia.ac.cn.

Abstract

Pre-mRNA splicing is an essential step in gene expression in most eukaryote genes. Here we present the feasibility of a genetically encoded luciferase reporter to monitor the pre-mRNA splicing process in living cells and animals. We showed that the splicing activity change induced by isoginkgetin could be readily visualized in vitro both in a dose and time dependent manner. Moreover, the pre-mRNA splicing process could be also obviously detected in mice by bioluminescence imaging and confirmed by RT-PCR. Our work provided a reporter system that allows high-throughput screening of chemical libraries to identify potential compounds leading to aberrant patterns of splicing.

Keywords: (170.0170) Medical optics and biotechnology; (170.3880) Medical and biological imaging; (170.6280) Spectroscopy, fluorescence and luminescence.