Multicistronic IVT mRNA for simultaneous expression of multiple fluorescent proteins
- 주제(키워드) 2A peptides , IVT mRNA , Multiple protein expression
- 등재 SCIE, SCOPUS, KCI등재
- 발행기관 Korean Society of Industrial Engineering Chemistry
- 발행년도 2019
- 총서유형 Journal
- URI http://www.dcollection.net/handler/ewha/000000162423
- 본문언어 영어
- Published As http://dx.doi.org/10.1016/j.jiec.2019.06.042
- 저작권 이화여자대학교 논문은 저작권에 의해 보호받습니다.
초록/요약
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. © 2019 The Korean Society of Industrial and Engineering Chemistry
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