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

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Bioluminescence

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2524))

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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.

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Acknowledgments

This work was supported by Natural Science Basic Research Program of Shaanxi (Program No. 2022JC-56) and the National Natural Science Foundation of China (81772010).

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Authors and Affiliations

  1. Engineering Research Center of Molecular and Neuro Imaging, Ministry of Education, School of Life Science and Technology, Xidian University, Xi’an, China

    Fu Wang, Si Chen, Haifeng Zheng & Bin Guo

  2. Institute of Medical Engineering, School of Basic Medical Sciences, Xi’an Jiaotong University, Xi’an, China

    Fu Wang

  3. Key Laboratory of Environment and Genes Related to Diseases, Ministry of Education of China, Xi’an Jiaotong University, Xi’an, China

    Fu Wang

Authors
  1. Fu Wang
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  2. Si Chen
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  3. Haifeng Zheng
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  4. Bin Guo
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Corresponding author

Correspondence to Fu Wang .

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Editors and Affiliations

  1. Environmental Management Research Institute, National Institute of Advanced Industrial Science and Technology (AIST), Tsukuba, Ibaraki, Japan

    Sung-Bae Kim

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© 2022 The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature

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Wang, F., Chen, S., Zheng, H., Guo, B. (2022). Design of an Intron-Retained Bioluminescence Reporter and its Application in Imaging of Pre-mRNA Splicing in Living Subjects. In: Kim, SB. (eds) Bioluminescence. Methods in Molecular Biology, vol 2524. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-2453-1_16

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  • DOI: https://doi.org/10.1007/978-1-0716-2453-1_16

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-2452-4

  • Online ISBN: 978-1-0716-2453-1

  • eBook Packages: Springer Protocols

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