Inflammation conditional genome editing mediated by the CRISPR-Cas9 system

Tingting Yuan; Honglin Tang; Xiaojie Xu; Jingjing Shao; Gaojun Wu; Young-Chang Cho; Yuan Ping; Guang Liang

doi:10.1016/j.isci.2023.106872

Inflammation conditional genome editing mediated by the CRISPR-Cas9 system

iScience. 2023 May 13;26(6):106872. doi: 10.1016/j.isci.2023.106872. eCollection 2023 Jun 16.

Authors

Tingting Yuan^{1

2

3

4}, Honglin Tang⁵, Xiaojie Xu², Jingjing Shao¹, Gaojun Wu¹, Young-Chang Cho³, Yuan Ping², Guang Liang^{1

4}

Affiliations

¹ Department of Cardiology, The First Affiliated Hospital of Wenzhou Medical University, Wenzhou, Zhejiang 325035, China.
² College of Pharmaceutical Sciences, Zhejiang University, Hangzhou, 310058, China.
³ Pharmaceutical Sciences, College of Pharmacy, Chonnam National University, Gwangju, Korea.
⁴ Chemical Biology Research Center, School of Pharmaceutical Sciences, Wenzhou Medical University, Wenzhou, Zhejiang 325035, China.
⁵ Department of Medical Oncology, Sir Run Run Shaw Hospital School of Medicine, Zhejiang University, Hangzhou, 310058, China.

Abstract

The specificity of CRISPR-Cas9 in response to particular pathological stimuli remains largely unexplored. Hence, we designed an inflammation-inducible CRISPR-Cas9 system by grafting a sequence that binds with NF-κB to the CRISPR-Cas9 framework, termed NBS-CRISPR. The genetic scissor function of this developed genome-editing tool is activated on encountering an inflammatory attack and is inactivated or minimized in non-inflammation conditions. Furthermore, we employed this platform to reverse inflammatory conditions by targeting the MyD88 gene, a crucial player in the NF-κB signaling pathway, and achieved impressive therapeutic effects. Finally, during inflammation, P65 (RELA) can translocate to the nucleus from the cytoplasm. Herein, to avoid Cas9 leaky DNA cleavage activity i, we constructed an NBS-P65-CRISPR system expressing the Cas9-p65 fusion protein. Our inflammation inducible Cas9-mediated genome editing strategy provides new perspectives and avenues for pathological gene interrogation.

Keywords: Biotechnology; Components of the immune system; Immune response.