Preparation of luciferase-expressing mRNA and expression characteristics of mRNA delivered by electroporation <i>in vivo</i>.

Lingjiang Fan; Keru Zhou; Yanguang Liu; Guiqin Wang; Ting Shi; Yihong HU; Daixi LI

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Preparation of luciferase-expressing mRNA and expression characteristics of mRNA delivered by electroporation in vivo.

Author: Lingjiang FAN ¹ ; Keru ZHOU ² ; Yanguang LIU ² ; Guiqin WANG ² ; Ting SHI ³ ; Yihong HU ⁴ ; Daixi LI ¹
Author Information

1. School of Health Science and Engineering, University of Shanghai for Science and Technology, Shanghai 200093, China.
2. Nanjing Advanced Academy of Life and Health, Nanjing 211135, Jiangsu, China.
3. School of Life Sciences and Biotechnology, Shanghai Jiao Tong University, Shanghai 200240, China.
4. Institut Pasteur of Shanghai, Chinese Academy of Sciences, Shanghai 200031, China.
Publication Type:Journal Article
Keywords: electroporation; in vivo imaging; luciferase; mRNA
MeSH: Animals; Electroporation/methods*; Gene Transfer Techniques; Luciferases/metabolism*; Mice; RNA, Messenger/genetics*
From: Chinese Journal of Biotechnology 2022;38(9):3379-3389
CountryChina
Language:Chinese
Abstract: In this study, we aimed to construct a non-replication mRNA platform and explore the side effects of electroporation-mediated delivery of mRNA on the mice as well as the expression features of the mRNA. With luciferase gene as a marker, in vitro transcription with T7 RNA polymerase was carried out for the synthesis of luciferase-expressed mRNA, followed by enzymatic capping and tailing. The mRNA was delivered in vivo by electroporation via an in vivo gene delivery system, and the expression intensity and duration of luciferase in mice were observed via an in vivo imaging system. The results demonstrated that the mRNA transcripts were successfully expressed both in vitro and in vivo. The electroporation-mediated delivery of mRNA had no obvious side effects on the mice. Luciferase was expressed successfully in all the mRNA-transduced mice, while the expression intensity and duration varied among individuals. Overall, the expression level peaked on the first day after electroporation and rapidly declined on the fourth day. This study is of great importance for the construction of non-replication mRNAs and their application in vaccine or antitumor drug development.