Split-Cas9-based targeted gene editing and nanobody-mediated proteolysis-targeting chimeras - optogenetically coordinated regulation of Survivin to control the fate of cancer cells

Sep 24, 2026

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Precise regulation of critical anti-apoptotic proteins in cancer is a key therapeutic and mechanistic strategy, yet achieving accurate, controllable targeting remains difficult. CRISPR/Cas9 editing and PROTACs help regulate gene expression and protein function, but their targeting accuracy and controllability require improvement. This study builds a coordinated photoactivatable platform combining split-Cas9 gene editing and light-induced protein degradation, enabling blue-light control of Survivin in nucleus and cytoplasm.

LanguageEnglish

CategoryHealthcare

FormatPDF

Pages16

Reading Time40 min

AccessFree to download

Graphical Abstract

Photoactivatable paProtacL and paCas9 systems

Abstract

Background

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DOI: 10.1002/ctm2.1382 RESEARCH ARTICLE Split-Cas9-based targeted gene editing andnanobody-mediated proteolysis-targeting chimeras optogenetically coordinated regulation of Survivin to control the fate of cancer cells Changping Deng1 Wenyun Zheng2 Shihui Li1 Yuping Liu2 Wen Bao2 Meiyan Wang3 Xingyuan Ma1 Chengnan Xu2 1 State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai, P. R. China 2 Shanghai Key Laboratory of New Drug Design, School of Pharmacy, East China University of Science and Technology, Shanghai, P. R. China 3 Synthetic Biology and Biomedical Engineering Laboratory, Biomedical Synthetic Biology Research Center, Shanghai Key Laboratory of Regulatory Biology, Institute of Biomedical, Sciences and School of Life Sciences, East China Normal University, Shanghai, P. R. China Correspondence Xingyuan Ma, State Key Laboratory of Bioreactor Engineering. Wenyun Zheng, Shanghai Key Laboratory of New Drug Design. East China University of Science and Technology, 130 Meilong Road, Shanghai, 200237, P. R. China. [Email:](Email: maxy@ecust.edu.cn)[ maxy@ecust.edu.cn](Email: maxy@ecust.edu.cn); [zwy@ecust.edu.cn](zwy@ecust.edu.cn) Graphical Abstract ∙ The photoactivatable PROTACs-Like (paProtacL) target protein via nanobodies and can controllably induce degradation of POI (protein of interest) under blue light. ∙ The photoactivatable Cas9 (paCas9) system can edit GOI (gene of interest) in the nucleus in a controlled manner under blue light irradiation. ∙ When paProtacL is combined with the paCas9 system, it can minimize the expression of the target protein. Clin. Transl. Med. 2023;13:e1382 . [https://doi.org/10.1002/ctm2.1382](https://doi.org/10.1002/ctm2.1382) [wileyonlinelibrary.com/journal/ctm2](wileyonlinelibrary.com/journal/ctm2) DOI: 10.1002/ctm2.1382 RESEARCH ARTICLE Split-Cas9-based targeted gene editing andnanobody-mediated proteolysis-targeting chimeras optogenetically coordinated regulation of Survivin to control the fate of cancer cells Changping Deng1 Wenyun Zheng2 Shihui Li1 Yuping Liu2 Wen Bao2 Meiyan Wang3 Xingyuan Ma1 Chengnan Xu2 1 State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai, P. R. China 2 Shanghai Key Laboratory of New Drug Design, School of Pharmacy, East China University of Science and Technology, Shanghai, P. R. China 3 Synthetic Biology and Biomedical Engineering Laboratory, Biomedical Synthetic Biology Research Center, Shanghai Key Laboratory of Regulatory Biology, Institute of Biomedical, Sciences and School of Life Sciences, East China Normal University, Shanghai, P. R. China Correspondence Xingyuan Ma, State Key Laboratory of Bioreactor Engineering. Wenyun Zheng, Shanghai Key Laboratory of New Drug Design. East China University of Science and Technology, 130 Meilong Road, Shanghai, 200237, P. R. China. [Email:](Email: maxy@ecust.edu.cn)[ maxy@ecust.edu.cn](Email: maxy@ecust.edu.cn); [zwy@ecust.edu.cn](zwy@ecust.edu.cn) Funding information National Key Research and Development Project of China, Grant/Award Number: 2018YFA0902804; National Natural Science Foundation, Grant/Award Numbers: 31670944, 81673345, 31870861; Science and Technology Innovation Action Plan of Shanghai, Grant/Award Number: 17431904600 Abstract Background: Precise regulation of partial critical proteins in cancer cells, such as anti-apoptotic proteins, is one of the crucial strategies for treating cancer and discovering related molecular mechanisms. Still, it is also challenging in actual research and practice. The widely used CRISPR/Cas9-based gene editing technology and proteolysis-targeting chimeras (PROTACs) have played an essential role in regulating gene expression and protein function in cells. However, the accuracy and controllability of their targeting remain necessary. Methods: Construction of UMUC-3-EGFP stable transgenic cell lines using

FAQs

What coordinated platform is developed to regulate Survivin?

The study establishes a synergistic control platform that combines photoactivatable split-Cas9 targeted gene editing with light-induced protein degradation, coordinating nuclear editing and cytoplasmic degradation of Survivin.

How is gene editing controlled in the proposed approach?

The Survivin gene in the nucleus is edited in a controlled manner using the photoactivatable split-Cas9 system under blue light irradiation (paCas9-Survivin).

How is Survivin protein degradation achieved and what is the experimental rationale?

Survivin protein in the cytoplasm is degraded precisely using a nanobody-mediated target (paProtacL-Survivin) under blue light. In vitro assays show that reducing Survivin promotes apoptosis and decreases cancer cell proliferation and migration, with CDX models supporting the in vivo impact.

Sep 24, 2026

Uploaded by Putri