Improvement of Tol2 Transposon System by Modification of Tol2 Transposase

被引:0
|
作者
Park, Ji Yun [1 ]
Lee, Haneur [1 ]
Song, Eun Seon [1 ]
Lee, Yun Haeng [1 ]
Kuk, Myeong Uk [1 ]
Ko, Gahyun [1 ]
Byun, Youngjoo [2 ]
Kwon, Hyung Wook [1 ,3 ]
Park, Joon Tae [1 ,3 ]
机构
[1] Incheon Natl Univ, Coll Life Sci & Bioengn, Div Life Sci, Incheon 22012, South Korea
[2] Korea Univ, Coll Pharm, Sejong 30019, South Korea
[3] Incheon Natl Univ, Convergence Res Ctr Insect Vectors, Incheon 22012, South Korea
基金
新加坡国家研究基金会;
关键词
Tol2; transposase; transposon; importin beta; SLEEPING-BEAUTY; GERMLINE TRANSGENESIS; MICROINJECTION;
D O I
10.1007/s12257-022-0175-9
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
Tol2 transposon-based vector platform has been developed as a useful strategy to improve protein yield. Tol2 transposase identifies inverted terminal repeats of transposons and enhances transgene integration efficiency in the genome. However, Tol2 transposon system does not reach a level of efficiency for use in commercial production, limiting its usefulness for mass production of recombinant proteins. Here, we proposed a novel strategy to improve the existing Tol2 transposase by adding a nuclear localization signal of histone H2B next to the Tol2 transposase (i.e., NLS-H2B-Tol2 transposase). Comparing the NLS-H2B-Tol2 transposase with the traditional Tol2 transposase, a significant increase in protein production was measured. Concurrently, a significant increase in nuclear transport of NLS-H2B-Tol2 transposase was observed, suggesting a causal relationship between increased nuclear transport and increased protein production. Furthermore, importin beta maximized NLS-H2B-Tol2-mediated protein productivity. Taken together, our findings offer useful ways for improving conventional Tol2 transposon-based vector platform. This new platform will be a breakthrough in the field of biopharmaceuticals to produce therapeutic proteins.
引用
收藏
页码:987 / 994
页数:8
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