Exploring the ability of TAT, a cell-penetrating peptide, to deliver proteins in a non-fused form
10.16438/j.0513-4870.2023-0266
- VernacularTitle:细胞穿膜肽TAT以非融合形式递送蛋白质的能力探究
- Author:
Jia DOU
1
;
Li-na JI
1
;
Zi-chun HUA
2
Author Information
1. State Key Laboratory of Pharmaceutical Biotechnology, School of Life Science, Nanjing University, Nanjing 210023, China
2. State Key Laboratory of Pharmaceutical Biotechnology, School of Life Science, Nanjing University, Nanjing 210023, China; Institute of Pharmaceutical Biotechnology of Jiangsu Industrial Technology Research Institute and Changzhou High-Tech Research Institute of Nanjing University, Changzhou 213164, China; Nanjing Genrecom Laboratories, Nanjing 210044, China
- Publication Type:Research Article
- Keywords:
cell-penetrating peptide;
TAT;
non-fusion expression;
elivery strategy;
TAT-EGFP;
cell permeability
- From:
Acta Pharmaceutica Sinica
2023;58(8):2384-2390
- CountryChina
- Language:Chinese
-
Abstract:
Accumulating evidence has shown that the cell-penetrating peptide TAT can be applied to deliver different types of drug molecules, including nucleic acids, proteins and small molecule drugs. Usually TAT delivers cargoes on the basis of their covalent bonds or non-covalent interactions. However, there are few reports on the delivery of proteins by TAT in a non-covalent manner, and no quantitative comparisons have been made on the protein delivery ability of TAT in fusion and non-fusion manners. In order to explore the ability of TAT to deliver proteins in non-fusion manner, here we used fluorescence microscopy and flow cytometry to investigate the ability of TAT to deliver enhanced green fluorescent protein (EGFP) into non-small cell lung cancer cells A549 in a non-fusion manner. It was found that TAT could deliver EGFP into A549 cells, and its delivery ability was positively correlated with its concentration. In addition, the fusion protein TAT-EGFP was overexpressed and purified, and its permeability across cell membrane was also investigated. In this paper, based on quantitative comparison, we found that the delivery of EGFP by TAT in fusion manner is significantly efficient than that of TAT in non-fusion manner. This is the report that TAT can deliver EGFP in a non-fusion manner. Although its delivery efficiency remains to be improved as compared with the fusion manner, the non-fusion manner has shown incomparable advantages in ease of operation, suggesting that it is also a candidate for delivery strategy in the future.
