Fluorescence protein conjugated nanodiamond-mediated CRISPR/Cas9 genome editing delivery system development and application in biomedical system

Chia-Ching Chang^1,2,3*, Chia-Yu Chang¹, Tien-Chun Yang⁴, Shih-Hwa Chiou^4,5, 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