Soluble expression and stability enhancement of transcription factors using 30Kc19 cell-penetrating protein

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• 作者：Jina Ryu ; Hee Ho Park ; Ju Hyun Park ; Hong Jai Lee…
• 关键词：30Kc19 protein ; Cell ; penetrating property ; Recombinant protein ; Transcription factor ; Soluble expression ; Protein stability
• 刊名：Applied Microbiology and Biotechnology
• 出版年：2016
• 出版时间：April 2016
• 年：2016
• 卷：100
• 期：8
• 页码：3523-3532
• 全文大小：762 KB
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• 作者单位：Jina Ryu (1)
  Hee Ho Park (2)
  Ju Hyun Park (3)
  Hong Jai Lee (2)
  Won Jong Rhee (4)
  Tai Hyun Park (1) (2) (5)
  
  1. Interdisciplinary Program for Bioengineering, Seoul National University, Seoul, 151-744, Republic of Korea
  2. The School of Chemical and Biological Engineering, Seoul National University, Seoul, 151-744, Republic of Korea
  3. Department of Medical Biomaterials Engineering, Kangwon National University, Chuncheon, 200-701, Republic of Korea
  4. Division of Bioengineering, Incheon National University, Incheon, 406-772, Republic of Korea
  5. Advanced Institutes of Convergence Technology, Suwon, 443-270, Republic of Korea
  
• 刊物类别：Chemistry and Materials Science
• 刊物主题：Chemistry
  Biotechnology
  Microbiology
  Microbial Genetics and Genomics
  
• 出版者：Springer Berlin / Heidelberg
• ISSN：1432-0614

文摘

Transcription factors have been studied as an important drug candidate. Ever since the successful generation of induced pluripotent stem cells (iPSCs), there has been tremendous interest in reprogramming transcription factors. Because of the safety risks involved in a virus-based approach, many researchers have been trying to deliver transcription factors using nonintegrating materials. Thus, delivery of transcription factors produced as recombinant proteins in E. coli was proposed as an alternative method. However, the low level of soluble expression and instability of such recombinant proteins are potential barriers. We engineered a Bombyx mori 30Kc19 protein as a fusion partner for transcription factors to overcome those problems. We have previously reported that 30Kc19 protein can be produced as a soluble form in E. coli and has a cell-penetrating property and a protein-stabilizing effect. Transcription factors fused with 30Kc19 (Oct4-30Kc19, Sox2-30Kc19, c-Myc-30Kc19, L-Myc-30Kc19, and Klf4-30Kc19) were produced as recombinant proteins. Interestingly, Oct4 and L-Myc were expressed as a soluble form by conjugating with 30Kc19 protein, whereas Oct4 alone and L-Myc alone aggregated. The 30Kc19 protein also enhanced the stability of transcription factors both in vitro and in cells. In addition, 30Kc19-conjugated transcription factors showed rapid delivery into cells and transcriptional activity significantly increased. Overall, 30Kc19 protein conjugation simultaneously enhanced soluble expression, stability, and transcriptional activity of transcription factors. We propose that the conjugation with 30Kc19 protein is a novel approach to solve the technical bottleneck of gene regulation using transcription factors.