Citation: Yuzhen Wang, Wenxiu Long, Bangshun He, Mengqiu Xiong, Jingjing Li, Yanfeng Gao. A dual-signal amplification strategy combining DNA-functionalized covalent organic frameworks and CRISPR-Cas12a for early detection of ovarian cancer biomarkers[J]. Chinese Chemical Letters, ;2026, 37(6): 112061. doi: 10.1016/j.cclet.2025.112061 shu

A dual-signal amplification strategy combining DNA-functionalized covalent organic frameworks and CRISPR-Cas12a for early detection of ovarian cancer biomarkers

Yuzhen Wang ^a ,
Wenxiu Long ^b ,
Bangshun He ^c ,
Mengqiu Xiong ^c ,
Jingjing Li ^{c, *,} ,
Yanfeng Gao ^{d, *,}

a.
Key Laboratory of Flexible Electronics (KLOFE), Institute of Advanced Materials (IAM) & School of Flexible Electronics (Future Technologies), Nanjing Tech University, Nanjing 211816, China
b.
College of Chemistry and Molecular Sciences, Wuhan University, Wuhan 430072, China
c.
Department of Laboratory Medicine, Nanjing First Hospital, Nanjing Medical University, Nanjing 210006, China
d.
School of Medical Imaging, Wannan Medical College, Wuhu 241002, China

lijingjing_li@163.com

gaoyanfeng@wnmc.edu.cn

Received Date: 25 March 2025
Revised Date: 8 September 2025
Accepted Date: 3 November 2025
Available Online: 4 November 2025

Figures(5)

Ovarian cancer is among the deadliest cancers, with the highest mortality rate among gynecological cancers; thus, highly sensitive early diagnosis is of paramount importance. In this study, we developed an innovative biosensor that integrates DNA-functionalized covalent organic frameworks with CRISPR-Cas12a technology for dual-signal amplification to achieve ultra-sensitive detection of ovarian cancer biomarkers. The COFs were co-functionalized with aptamers specific to target protein biomarkers and a larger number of activators. Upon recognition and binding to target proteins, the activators anchored on the COF surface become accessible in the detection system, converting low-abundance protein signals into amplified nucleic acid signals, which represents the first stage of signal amplification. These activators subsequently trigger the trans-cleavage activity of CRISPR-Cas12a, leading to the cleavage of fluorophore–quencher reporters and resulting in further amplified fluorescence output, constituting the second stage of amplification. This dual-signal amplification strategy, integrated with a microfluidic chip, enabled the sensitive detection of CA125 and HE4, with detection limits as low as 0.001 U/mL and 0.2 pg/mL, respectively, providing a robust, accurate, and scalable platform for ovarian cancer diagnostics and demonstrating potential applications in clinical bioanalysis and diagnosis.
- Microfluidics,
- Covalent organic frameworks,
- CRISPR-Cas12a,
- Signal amplification,
- CA125,
- HE4
1. [1]
  P.M. Webb, S.J. Jordan, Nat. Rev. Clin. Oncol. 21 (2024) 389–400. doi: 10.1038/s41571-024-00881-3
2. [2]
  J.M. Liberto, S.Y. Chen, I.M. Shih, et al., Cancers 14 (2022) 2885. doi: 10.3390/cancers14122885
3. [3]
  D. Zhou, Z. Zhang, L. Pan, et al., Angew. Chem. Int. Ed. 63 (2024) e202404493. doi: 10.1002/anie.202404493
4. [4]
  M. Kim, C. Chen, P. Wang, et al., Nat. Biomed. Eng. 6 (2022) 267–275. doi: 10.1038/s41551-022-00860-y
5. [5]
  J. Yang, B. Pan, F. Zeng, et al., Nano Lett. 21 (2021) 2001–2009. doi: 10.1021/acs.nanolett.0c04476
6. [6]
  Y. Wang, Y. Gao, Y. Song, ChemMedChem 17 (2022) e202200422. doi: 10.1002/cmdc.202200422
7. [7]
  S.F. Tseng, L.Y. Chiu, S.H. Hsu, C.C. Kuo, Sens. Actuators B 422 (2025) 136569. doi: 10.1016/j.snb.2024.136569
8. [8]
  Q. Zhao, B. Pan, W.X. Long, et al., Nano Lett. 22 (2022) 9714–9722. doi: 10.1021/acs.nanolett.2c04022
9. [9]
  Q. Zhao, Y. Pan, X. Luan, et al., Biosens. Bioelectron. 190 (2021) 113450. doi: 10.1016/j.bios.2021.113450
10. [10]
  Y. Pan, X. Luan, F. Zeng, et al., Biosens. Bioelectron. 209 (2022) 114239. doi: 10.1016/j.bios.2022.114239
11. [11]
  T. Zhang, W. Zhao, X. Chen, et al., Anal. Chem. 94 (2022) 15472–15480. doi: 10.1021/acs.analchem.2c03607
12. [12]
  M. Zhang, H. Wang, H. Wang, et al., Anal. Chem. 93 (2021) 7942–7948. doi: 10.1021/acs.analchem.1c00686
13. [13]
  J. Yan, Z. Xu, H. Zhou, et al., Anal. Chem. 94 (2022) 16481–16490. doi: 10.1021/acs.analchem.2c04079
14. [14]
  Y. Wang, Y. Gao, Y. Yin, et al., Microchim. Acta 189 (2022) 139.
15. [15]
  Y. Gao, Y. Wang, X. Liu, et al., Anal. Chim. Acta 1241 (2023) 340806. doi: 10.1016/j.aca.2023.340806
16. [16]
  Y. Gao, Y. Wang, Y. Wang, et al., TrAC, Trends Anal. Chem. 158 (2023) 116887.
17. [17]
  Y. Wang, W. Long, Y. Wang, et al., Chem. Eng. J. 503 (2025) 158289. doi: 10.1016/j.cej.2024.158289
18. [18]
  J. Li, W. Bi, Y. Gao, et al., Sens. Actuators B 408 (2024) 135526.
19. [19]
  Y. Gao, D. Zhou, Q. Xu, et al., ACS Appl. Mater. Interfaces 15 (2023) 5010–5018.
20. [20]
  Y. Gao, Y. Wang, B. He, et al., Angew. Chem. Int. Ed. 62 (2023) e202302000.
21. [21]
  D. Ouyang, Z. Fu, G. Li, et al., Chin. Chem. Lett. 34 (2023) 107992.
22. [22]
  D. Ouyang, C. Wang, C. Zhong, et al., Chem. Sci. 15 (2024) 278–284.
23. [23]
  A. Wang, X. Liu, S. Feng, et al., ChemBioChem 26 (2024) e202400807.
24. [24]
  M. Sun, Z. Liu, L. Wu, et al., J. Am. Chem. Soc. 145 (2023) 5330–5341.
25. [25]
  K. Geng, T. He, R. Liu, et al., Chem. Rev. 120 (2020) 8814–8933.
26. [26]
  Y. Shi, J. Yang, F. Gao, Q. Zhang, ACS Nano 17 (2023) 1879–1905.
27. [27]
  G. Li, Y. Wu, C. Zhong, et al., Chin. Chem. Lett. 35 (2024) 108904.
1. [1]
  Hao Hu , Zhengxin Ye , Lei Zhang , Kejun Dong , Bei Yan , Longjie Li , Wei Zhang , Xianjin Xiao . A PAM-free and universal CRISPR-Cas12a activation model for ultra-sensitive DNA methylation detection. Chinese Chemical Letters, 2026, 37(1): 111184-. doi: 10.1016/j.cclet.2025.111184
2. [2]
  Yu-Qin Xia , Shui-Ming Jing , Li-Mei Chang , Zhi-Gang Gu , Jian Zhang . Luminescent covalent organic frameworks: Classification to optical applications. Chinese Journal of Structural Chemistry, 2026, 45(1): 100731-100731. doi: 10.1016/j.cjsc.2025.100731
3. [3]
  Jiaqi Ma , Lan Li , Yiming Zhang , Jinjie Qian , Xusheng Wang . Covalent organic frameworks: Synthesis, structures, characterizations and progress of photocatalytic reduction of CO2. Chinese Journal of Structural Chemistry, 2024, 43(12): 100466-100466. doi: 10.1016/j.cjsc.2024.100466
4. [4]
  Xinrui Chen , Wenjian Huang , Xiaoyang Zhao , Songyao Zhang , Xinrui Miao . Structural regulation of alkynyl-based covalent organic frameworks for multi-stimulus fluorescence sensing. Chinese Journal of Structural Chemistry, 2025, 44(7): 100604-100604. doi: 10.1016/j.cjsc.2025.100604
5. [5]
  Weixu Li , Yuexin Wang , Lin Li , Xinyi Huang , Mengdi Liu , Bo Gui , Xianjun Lang , Cheng Wang . Promoting energy transfer pathway in porphyrin-based sp2 carbon-conjugated covalent organic frameworks for selective photocatalytic oxidation of sulfide. Chinese Journal of Structural Chemistry, 2024, 43(7): 100299-100299. doi: 10.1016/j.cjsc.2024.100299
6. [6]
  Xueting Hu , Lijia Zhao , Tingting Liao , Cheng-Peng Li . Crystalline porous organic frameworks: Emerging platforms for enzyme immobilization in biomedical applications. Chinese Journal of Structural Chemistry, 2026, 45(2): 100793-100793. doi: 10.1016/j.cjsc.2025.100793
7. [7]
  Yuting Wu , Haifeng Lv , Xiaojun Wu . Design of two-dimensional porous covalent organic framework semiconductors for visible-light-driven overall water splitting: A theoretical perspective. Chinese Journal of Structural Chemistry, 2024, 43(11): 100375-100375. doi: 10.1016/j.cjsc.2024.100375
8. [8]
  Haijiang Gong , Qingtan Zeng , Shili Gai , Yaqian Du , Jing Zhang , Qingyu Wang , He Ding , Lichun Wu , Anees Ahmad Ansari , Piaoping Yang . Enzyme-based colorimetric signal amplification strategy in lateral flow immunoassay. Chinese Chemical Letters, 2025, 36(5): 110059-. doi: 10.1016/j.cclet.2024.110059
9. [9]
  Bo Liu , Shuaiqiang Shao , Junjie Cai , Zijian Zhang , Feng Tian , Kun Yang , Fan Li . Signal cascade amplification of streptavidin-biotin-modified immunofluorescence nanocapsules for ultrasensitive detection of glial fibrillary acidic protein. Chinese Chemical Letters, 2025, 36(3): 109814-. doi: 10.1016/j.cclet.2024.109814
10. [10]
  Shuo Tian , Shuyun Chen , Yunsen Wang , Dianping Tang . Liposomal photoelectrochemical immunoassay for low-abundance proteins with ternary transition metal sulfides for signal amplification. Chinese Chemical Letters, 2025, 36(7): 110418-. doi: 10.1016/j.cclet.2024.110418
11. [11]
  Deshuai Zhen , Chunlin Liu , Qiuhui Deng , Shaoqi Zhang , Ningman Yuan , Le Li , Yu Liu . A review of covalent organic frameworks for metal ion fluorescence sensing. Chinese Chemical Letters, 2024, 35(8): 109249-. doi: 10.1016/j.cclet.2023.109249
12. [12]
  Yujie Wang , Haoran Wang , Yanni Liu , Manhua Peng , Hongwei Fan , Hong Meng . A comprehensive review on the scalable and sustainable synthesis of covalent organic frameworks. Chinese Chemical Letters, 2025, 36(8): 110189-. doi: 10.1016/j.cclet.2024.110189
13. [13]
  Qi Li , Minqiao Liang , Huifen Zhuang , Zhengyang Chen , Yuxiang Jiang , Xiaofei Chen , Yifa Chen , Ya-Qian Lan . Underscoring the polyimide-linkage in covalent organic frameworks and related applications. Chinese Chemical Letters, 2026, 37(2): 111593-. doi: 10.1016/j.cclet.2025.111593
14. [14]
  Liying Ou , Zhenluan Xue , Bo Li , Zhiwei Jin , Jiaochan Zhong , Lixia Yang , Penghui Shao , Shenglian Luo . Nitrogen-containing linkage-bonds in covalent organic frameworks: Synthesis and applications. Chinese Chemical Letters, 2025, 36(6): 110294-. doi: 10.1016/j.cclet.2024.110294
15. [15]
  Bo Li , Yuanzhe Cheng , Xuyang Ma , Dongxu Zhao , Yang Zhang , Yongxing Sun , Jia Chen , Li Wu , Liang Zhao , Hongdeng Qiu , Yujian He . Facile and scale-up synthesis of cyano-functionalized covalent organic frameworks for selective gold recovery. Chinese Chemical Letters, 2026, 37(1): 111134-. doi: 10.1016/j.cclet.2025.111134
16. [16]
  Kun Zhao , Bing Chen , Luwen Liang , Yaze Chen , Shan Yao , Qun Peng , Yuhao Liu , Bin Han . Microenvironment engineering of covalent organic frameworks for hydrogen peroxide synthesis via photocatalytic oxygen reduction. Chinese Chemical Letters, 2026, 37(6): 112032-. doi: 10.1016/j.cclet.2025.112032
17. [17]
  Zihui Liu , Xiaolin Liu , Lin Tang , Houhe Pan , Xi Liu , Jianzhuang Jiang . Covalent organic frameworks-based heterojunctions for photocatalytic hydrogen peroxide production and in-situ application. Chinese Chemical Letters, 2026, 37(6): 112024-. doi: 10.1016/j.cclet.2025.112024
18. [18]
  Qiang Fang , Yingbo Lu , Jianying Huang , Cheng Zhang , Jing Wu , Shijun Li . An electrochemical immunosensor based on an antibody-ferrocene-functionalized covalent organic framework. Chinese Chemical Letters, 2026, 37(2): 111218-. doi: 10.1016/j.cclet.2025.111218
19. [19]
  Shiyan Ai , Yaning Xu , Hui Zhou , Ziwei Cui , Tiantian Wu , Dan Tian . Superelastic and ultralight covalent organic framework composite aerogels modified with different functional groups for ultrafast adsorbing organic pollutants in water. Chinese Chemical Letters, 2025, 36(10): 110761-. doi: 10.1016/j.cclet.2024.110761
20. [20]
  Guorong Li , Yijing Wu , Chao Zhong , Yixin Yang , Zian Lin . Predesigned covalent organic framework with sulfur coordination: Anchoring Au nanoparticles for sensitive colorimetric detection of Hg(Ⅱ). Chinese Chemical Letters, 2024, 35(5): 108904-. doi: 10.1016/j.cclet.2023.108904

Get Citation

PDF

XML

Metrics

PDF Downloads(0)
Abstract views(16)
HTML views(2)

通讯作者: 陈斌, bchen63@163.com

1.
沈阳化工大学材料科学与工程学院沈阳 110142