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Multicistronic IVT mRNA for simultaneous expression of multiple fluorescent proteins

Title
Multicistronic IVT mRNA for simultaneous expression of multiple fluorescent proteins
Authors
Lee, KyuriKim, Soo YoungSeo, YunmiKwon, HyokyungKwon, Young JikLee, Hyukjin
Ewha Authors
이혁진이규리
SCOPUS Author ID
이혁진scopus
Issue Date
2019
Journal Title
JOURNAL OF INDUSTRIAL AND ENGINEERING CHEMISTRY
ISSN
1226-086XJCR Link

1876-794XJCR Link
Citation
JOURNAL OF INDUSTRIAL AND ENGINEERING CHEMISTRY vol. 80, pp. 770 - 777
Keywords
IVT mRNA2A peptidesMultiple protein expression
Publisher
ELSEVIER SCIENCE INC
Indexed
SCIE; SCOPUS; KCI WOS scopus
Document Type
Article
Abstract
Synthetic mRNA has emerged as a promising gene expression system for more rapid and controllable induction of target proteins, due to no need of transnuclear localization. It can efficiently generate large amount of encoded proteins in cells as compared to that of conventional pDNA. In addition, the development of in vitro transcription (IVT) technique has enabled the facile modulation of synthetic mRNA's biochemical properties, further strengthening its applications in pharmaceutical sciences and engineering. In this study, we have investigated the IVT mRNA for simultaneous expression of three distinctive fluorescent (GFP, RFP, and BFP) proteins. To achieve simultaneous expression of the multiple proteins by a single platform, 2A peptides encoding sequences were used in the design of GRB mRNA. Self-cleavages of 2A peptides in the translated fusion proteins were confirmed by western blot analysis. The expression level of the IVT mRNA was optimized by evaluating the use of chemical modification, the method of poly A tailing, and the capping conditions of IVT process. (C) 2019 The Korean Society of Industrial and Engineering Chemistry. Published by Elsevier B.V. All rights reserved.
DOI
10.1016/j.jiec.2019.06.042
Appears in Collections:
약학대학 > 약학과 > Journal papers
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