Fluorescence protein conjugated nanodiamond-mediated CRISPR/Cas9 genome editing delivery system development and application in biomedical system
Chia-Ching Chang1,2,3*, Chia-Yu Chang1, Tien-Chun Yang4, Shih-Hwa Chiou4,5, Yonhua Tzeng6
1Biological Science and Technology, National Chiao Tung University, Hsinchu, Taiwan
2Electrophysics, National Chiao Tung University, Hsinchu, Taiwan
3Institute of Physics, Academia Sinica, Taipei, Taiwan
4Medical Research, Taipei Veterans General Hospital, Taipei, Taiwan
5School of Medicine, National Yang-Ming University, Taipei, Taiwan
6Electrical Engineering, National Cheng Kung University, Tainan, Taiwan
* Presenter:Chia-Ching Chang, email:ccchang01@faculty.nctu.edu.tw
CRISPR/Cas9 system is a powerful genome editing tool, its delivery efficiency is low (~10% in average). Therefore, a high efficient delivery system is highly desired. Nano scale diamond particles (NDs) are bio-compatible materials which have been used for DNA, proteins or chemical compounds delivery into cells and bio-tissues. In this study, fluorescent mCherry conjugated NDs were used as the mediator to deliver RS1 gene editing CRISPR/Cas9 system into human iPSCs and mouse retinas. The efficiency of cell internalization was significantly improved. The genome editing efficiency is as high as ~19.3% of all alleles of human iPSCs. Furthermore, the RS1 gene mutation can be observed in mouse retinas, too. In summary, this our fluorescence traceable gene editing system can be used for creating in vitro and in vivo disease models of XLRS. Furthermore, this ND-mediated genome editing delivery system may apply in other therapeutic approaches, too.

Keywords: Nanodiamond, CrisPR/Cas9, bioconjugation, self-monitor, high efficiency delivery