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

• Published in: Biomaterials Vol. 173; pp. 11 - 21
• Main Authors: Takashina, Tomoki, Koyama, Takayoshi, Nohara, Satoshi, Hasegawa, Masakatsu, Ishiguro, Akira, Iijima, Kenta, Lu, Jun, Shimura, Mari, Okamura, Tadashi, Sakuma, Tetsushi, Yamamoto, Takashi, Ishizaka, Yukihito
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier Ltd 01.08.2018
• Subjects: Artificial transcription factor; Cell-penetrating peptide (CPP); Induced pluripotent stem cell (iPSC); Mouse embryonic fibroblast (MEF); Transcription activator-like effector (TALE); Mouse embryonic fibroblast (MEF); Induced pluripotent stem cell (iPSC); Cell-penetrating peptide (CPP); Artificial transcription factor; Transcription activator-like effector (TALE)
• ISSN: 0142-9612; 1878-5905; 1878-5905
• DOI: 10.1016/j.biomaterials.2018.04.040

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.