Identification of a cell-penetrating peptide applicable to a protein-based transcription activator-like effector expression system for cell engineering. (August 2018)

• Record Type: Journal Article
• 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
• Is Part Of: Biomaterials. Volume 173(2018)
• Page Start: 11
• Page End: 21
• Publication Date: 2018-08
• Subjects: Cell-penetrating peptide (CPP) -- Artificial transcription factor -- Induced pluripotent stem cell (iPSC) -- Transcription activator-like effector (TALE) -- Mouse embryonic fibroblast (MEF); Biomedical materials -- Periodicals; Biocompatible Materials -- Periodicals; Biomatériaux -- Périodiques; 610.28
• DOI: 10.1016/j.biomaterials.2018.04.040
• Languages: English
• ISSNs: 0142-9612

Abstract

Abstract: 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.