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

Fu Wang; Si Chen; Haifeng Zheng; Bin Guo

doi:10.1007/978-1-0716-2453-1_16

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

Fu Wang
, Si Chen
, Haifeng Zheng
, Bin Guo

Health Science Center

Xidian University

Research output: Chapter in Book/Report/Conference proceeding › Chapter › peer-review

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.

Original language	English
Title of host publication	Methods in Molecular Biology
Publisher	Humana Press Inc.
Pages	209-221
Number of pages	13
DOIs	https://doi.org/10.1007/978-1-0716-2453-1_16
State	Published - 2022

Publication series

Name	Methods in Molecular Biology
Volume	2524
ISSN (Print)	1064-3745
ISSN (Electronic)	1940-6029

Keywords

Bioluminescence reporter
Imaging
Intron-retained transcript
Pre-mRNA splicing
Renilla luciferase

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

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10.1007/978-1-0716-2453-1_16

Cite this

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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.",

keywords = "Bioluminescence reporter, Imaging, Intron-retained transcript, Pre-mRNA splicing, Renilla luciferase",

author = "Fu Wang and Si Chen and Haifeng Zheng and Bin Guo",

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T1 - Design of an Intron-Retained Bioluminescence Reporter and its Application in Imaging of Pre-mRNA Splicing in Living Subjects

AU - Wang, Fu

AU - Chen, Si

AU - Zheng, Haifeng

AU - Guo, Bin

PY - 2022

Y1 - 2022

N2 - 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.

AB - 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.

KW - Bioluminescence reporter

KW - Imaging

KW - Intron-retained transcript

KW - Pre-mRNA splicing

KW - Renilla luciferase

UR - https://www.scopus.com/pages/publications/85133921131

U2 - 10.1007/978-1-0716-2453-1_16

DO - 10.1007/978-1-0716-2453-1_16

M3 - 章节

C2 - 35821474

AN - SCOPUS:85133921131

T3 - Methods in Molecular Biology

SP - 209

EP - 221

BT - Methods in Molecular Biology

PB - Humana Press Inc.

ER -

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

Abstract

Publication series

Keywords

UN SDGs

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