Identification of a cell-penetrating peptide applicable to a protein-based transcription activator-like effector expression system for cell engineering
| Autor: | Yukihito Ishizaka, Tomoki Takashina, Masakatsu Hasegawa, Tetsushi Sakuma, Takashi Yamamoto, Tadashi Okamura, Mari Shimura, Jun Lu, Takayoshi Koyama, Kenta Iijima, Satoshi Nohara, Akira Ishiguro |
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| Rok vydání: | 2018 |
| Předmět: |
0301 basic medicine
Cell Survival viruses Induced Pluripotent Stem Cells Biophysics Artificial transcription factor Active Transport Cell Nucleus Bioengineering Peptide Cell-Penetrating Peptides Biomaterials 03 medical and health sciences Mice Transcription (biology) Cell Line Tumor Animals Humans Induced pluripotent stem cell Cell Engineering Transcription Activator-Like Effectors chemistry.chemical_classification Cell Nucleus Expression vector Chemistry Effector Chimera Fibroblasts Cellular Reprogramming Recombinant Proteins Cell biology MicroRNAs 030104 developmental biology Mechanics of Materials Ceramics and Composites Cell-penetrating peptide Reprogramming Transcription Factors |
| Zdroj: | Biomaterials. 173 |
| ISSN: | 1878-5905 |
| Popis: | Cellular reprogramming is a promising technology in regenerative medicine, but most studies have been performed by using expression vectors. For future clinical applications, it is necessary to establish a system in which cell engineering can be manipulated without any risk of damaging the genome. Here, we identified a cell-penetrating peptide composed of 10 amino acids (RIFIHFRIGC) with nuclear trafficking activity and found that it was significantly more potent than a Tat-derived peptide or polyarginine peptide (R11). We named the peptide “nuclear trafficking peptide” (NTP) and applied it to a protein-based artificial transcription factor (NTP-ATF), which was composed of a transcription activator-like effector and transcription domain (VP64). An NTP-ATF designed to the proximal promoter region of the microRNA-302/367 cluster efficiently induced endogenous RNA expression at an extremely low concentration (0.25 nM), and repetitive treatment of mouse embryonic fibroblasts with NTP-ATF generated induced pluripotent stem-like cells, which gave chimeric mice. Together with the observation that recombinant NTP-ATF protein did not induce any apparent cytotoxicity, we propose that NTP-ATF is a promising system for cellular reprogramming applicable to regenerative medicine. |
| Databáze: | OpenAIRE |
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