An orthogonal CRISPR/Cpf1 platform for precise spatiotemporal gene regulation and osteoporotic fracture repair
Year: 2026
Type: article
Abstract: CRISPR-Cas systems enable powerful gene editing and regulation, yet single-modality control often fails to achieve orthogonal, spatiotemporally precise regulation of multiple endogenous genes. We engineered OREC, an orthogonal platform integrating chemogenetic and optogenetic modalities for precise, reversible, multiplex gene control. OREC comprises two components: OREC C regulated by doxycycline (Dox) and OREC o controlled by light. By assembling catalytically dead Cpf1 (dCpf1), gene regulatory elements, and crRNA arrays on single transcripts, OREC enables robust simultaneous manipulation of multiple genes. We demonstrated OREC's therapeutic potential in vitro for osteoblast function modulation and in vivo for osteoporotic fracture repair. OREC effectively activated Bmp2 while inhibiting Dkk1 , significantly enhancing bone formation and fracture healing in mouse models. These results establish OREC as a versatile platform for precise multiplex gene regulation, offering significant advancement for CRISPR-based gene therapy applications in complex tissues where coordinated control of multiple therapeutic targets is essential.
Source: Cell Reports Methods
Authors: Jie Zhao, Zengliang Wang, Lina Lu, Guoyun Bu, Zukang Miao, Yang Zhang, Yue Guo, Zhao Yang, Jianxiong Ma, Jun Jiao, Xinlong Ma
Institutions: Tianjin University, Tianjin Hospital, Nankai University
Language: English
Subfield: Molecular Biology
Domain: Life Sciences
Open Access status: gold
Estimated APC paid: $4,824
Funders: National Natural Science Foundation of China, Natural Science Foundation of Tianjin City, China Academy of Space Technology, National Key Research and Development Program of China, National Key Clinical Specialty Discipline Construction Program of China, Tianjin Science and Technology Program
OpenAlex ID: W7128602173