Biotechnology and Bioprocess Engineering 2022; 27(3): 353-360  
Improvement of Sleeping Beauty Transposon System Enabling Efficient and Stable Protein Production
Yun Haeng Lee, Ji Yun Park, Eun Seon Song, Haneur Lee, Myeong Uk Kuk, Junghyun Joo, Hyungmin Roh, and Joon Tae Park
Yun Haeng Lee, Ji Yun Park, Eun Seon Song, Haneur Lee, Myeong Uk Kuk, Junghyun Joo, Joon Tae Park*
Division of Life Sciences, College of Life Sciences and Bioengineering, Incheon National University, Incheon 22012, Korea
Tel: +82-32-835-8841; Fax: +82-32-835-0754
Joon Tae Park
Convergence Research Center for Insect Vectors, Incheon National University, Incheon 22012, Korea
Hyungmin Roh*
Department of Chemical and Environmental Technology, Inha Technical College, Incheon 22212, Korea
Tel: +82-32-870-2276; Fax: +82-32-870-2514
Received: August 18, 2021; Revised: December 17, 2021; Accepted: January 14, 2022; Published online: June 30, 2022.
© The Korean Society for Biotechnology and Bioengineering. All rights reserved.

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License ( which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.
The most essential goal in the field of biopharmaceuticals is to develop cell lines with higher protein yields. To this goal, the Sleeping Beauty (SB) transposon-based expression system has been developed as a powerful tool for increasing protein productivity. However, SB transposon system has fallen short of expectation in terms of the efficiency and stability of protein production, limiting its applicability to large-scale production of recombinant proteins. Here, we propose a novel strategy to increase the efficiency and stability of protein production, through modification of the traditional SB transposon vector. Adding a pair of inverted terminal repeats (ITRs) next to existing ITRs (i.e., double-ITRs) significantly increased the efficiency of transgene integration, resulting in high-yield and sustained protein production. Furthermore, double-ITRs responded more favorably to DNA methylation inhibitors in terms of protein yield, implying that using double-ITRs with DNA methylation inhibitors may be effective in increasing protein productivity. Taken together, our study introduces a new vector platform that is applicable to high-yield and sustained protein production, and will open new avenues in the field of biopharmaceuticals.
Keywords: Sleeping Beauty transposon, double inverted terminal repeats, protein productivity, DNA methylation inhibitor

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