Abstract
Precise activation of endogenous genes is a powerful strategy for functional genomics and therapeutic development, but current CRISPR-based transcriptional activation (CRISPRa) systems are limited by the large size of Cas proteins for adeno-associated virus (AAV) delivery. Here, we present a high-efficiency dCas12f-based transcriptional activation system (HEAL), which recruits transactivators through MS2 coat protein binding to MS2 aptamers embedded within the sgRNA scaffold. Engineered to enhance DNA binding, nuclear localization, and transactivator recruitment, HEAL induces over 100,000-fold activation of endogenous genes and outperforms existing CRISPRa systems in vitro and in vivo. We further develop red-light-inducible OptoHEAL and small-molecule-inducible ChemHE…
Abstract
Precise activation of endogenous genes is a powerful strategy for functional genomics and therapeutic development, but current CRISPR-based transcriptional activation (CRISPRa) systems are limited by the large size of Cas proteins for adeno-associated virus (AAV) delivery. Here, we present a high-efficiency dCas12f-based transcriptional activation system (HEAL), which recruits transactivators through MS2 coat protein binding to MS2 aptamers embedded within the sgRNA scaffold. Engineered to enhance DNA binding, nuclear localization, and transactivator recruitment, HEAL induces over 100,000-fold activation of endogenous genes and outperforms existing CRISPRa systems in vitro and in vivo. We further develop red-light-inducible OptoHEAL and small-molecule-inducible ChemHEAL for remote and precise transcriptional control. AAV-delivered HEAL targeting interleukin 10 alleviates acute kidney injury in mice, while ChemHEAL-mediated activation of thymic stromal lymphopoietin reduces body weight in obese mice. HEAL provides a modular, compact, and controllable platform for endogenous gene activation with strong potential for fundamental research and gene therapy.
Data availability
All data associated with this study are presented in the paper or the Supplementary Information. The data generated in this study are provided in the Source Data file. Sequences for plasmids used in the study are available in the Supplementary Data file. All genetic components related to this paper are available with a material transfer agreement and can be requested from H.Y. (hfye@bio.ecnu.edu.cn). Source data are provided in this paper.
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Acknowledgements
This work was financially supported by grants from the Noncommunicable Chronic Diseases-National Science and Technology Major Project (no. 2023ZD0501300), the Science and Technology Commission of Shanghai Municipality (nos. 23HC1410100, 25HC2830300), and the National Natural Science Foundation of China (nos. 32250010, 32261160373, 32430064), and the Fundamental Research Funds for the Central Universities to H.Y., and the Science and Technology Commission of Shanghai Municipality (nos. 25J22800100, 25ZR1402121) to N.G. This work was also partially supported by the China Postdoctoral Science Foundation (nos. 2025M772641, BX20250150) and the Science and Technology Commission of Shanghai Municipality (no. 24YF2735700) to D.K., the Young Scientists Fund of the National Natural Science Foundation of China (no. 32300458), the Science and Technology Commission of Shanghai Municipality (no. 23YF1410700), and the Natural Science Foundation of Chongqing (CSTB2023NSCQ-MSX0126) to Y.Z., the National Natural Science Foundation of China (no. 32301217) to J.Y., and the National Natural Science Foundation of China (no. 32300083) to Y.Y. H.Y. is a SANS Exploration Scholar. We are also thankful for support from the CAS Youth Interdisciplinary Team and the ECNU Multifunctional Platform for Innovation (011) for supporting mice experiments and the Instruments Sharing Platform of the School of Life Sciences, ECNU.
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Author notes
These authors contributed equally: Hang Wan, Deqiang Kong, Tao Yan.
Authors and Affiliations
Shanghai Key Laboratory of Regulatory Biology, Institute of Biomedical Sciences and School of Life Sciences, Shanghai Academy of Natural Sciences (SANS), East China Normal University, Shanghai, China
Hang Wan, Tao Yan, Yang Zhou, Mengyao Liu, Xiaoding Ma, Tianjie Zhao, Wenmin Zhou, Xingwan Liu, Jianli Yin, Ningzi Guan & Haifeng Ye 1.
Shanghai Fengxian District Central Hospital, Shanghai, China
Deqiang Kong 1.
Wuhu Hospital, Wuhu City, China
Yang Zhou
Authors
- Hang Wan
- Deqiang Kong
- Tao Yan
- Yang Zhou
- Mengyao Liu
- Xiaoding Ma
- Tianjie Zhao
- Wenmin Zhou
- Xingwan Liu
- Jianli Yin
- Ningzi Guan
- Haifeng Ye
Contributions
H.Y. conceived the project. H.Y., D.K., and H.W. designed the project. H.W., D.K., T.Y., Y.Z., M.L., X.M., T.Z., W.Z., X.L., and J.Y. performed the experimental work. H.Y., H.W., D.K., and N.G. analyzed the results and wrote the manuscript. All authors edited and approved the manuscript.
Corresponding authors
Correspondence to Ningzi Guan or Haifeng Ye.
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Competing interests
H.Y., H.W., and N.G. are inventors on patent applications (Chinese patent application number 2025118539236) submitted by ECNU. The remaining authors declare no competing interests.
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Cite this article
Wan, H., Kong, D., Yan, T. et al. A compact and inducible dCas12f-based CRISPRa platform for programmable in vivo gene activation. Nat Commun (2026). https://doi.org/10.1038/s41467-025-68183-5
Received: 16 July 2025
Accepted: 22 December 2025
Published: 08 January 2026
DOI: https://doi.org/10.1038/s41467-025-68183-5