Identification of a cell-penetrating peptide applicable to a protein-based transcription activator-like effector expression system for cell engineering

Publication date: August 2018
Source:Biomaterials, Volume 173
Author(s): Tomoki Takashina, Takayoshi Koyama, Satoshi Nohara, Masakatsu Hasegawa, Akira Ishiguro, Kenta Iijima, Jun Lu, Mari Shimura, Tadashi Okamura, Tetsushi Sakuma, Takashi Yamamoto, Yukihito Ishizaka
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.



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