Droplets Cas13a‐RPA measurement delineates potential role for plasma circWDR37 in colorectal cancer
Abstract Colorectal cancer (CRC) is one of the most prevalent forms of cancer. CircRNAs have emerged as promising biomarkers for cancer diagnosis and prognosis evaluation. However, novel circRNAs as potential biomarkers for CRC still need further exploration and validation, and precise detection met...
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2025-01-01
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| Online Access: | https://doi.org/10.1002/agt2.663 |
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| author | Jingsong Xu Li Cao Shuang Yang Ying Jian Yu Liu Zhen Shen Qian Liu Xiang Chen Min Li Shun Li Xiaolei Zuo Min Li Hua Wang |
| author_facet | Jingsong Xu Li Cao Shuang Yang Ying Jian Yu Liu Zhen Shen Qian Liu Xiang Chen Min Li Shun Li Xiaolei Zuo Min Li Hua Wang |
| author_sort | Jingsong Xu |
| collection | DOAJ |
| description | Abstract Colorectal cancer (CRC) is one of the most prevalent forms of cancer. CircRNAs have emerged as promising biomarkers for cancer diagnosis and prognosis evaluation. However, novel circRNAs as potential biomarkers for CRC still need further exploration and validation, and precise detection methods are yet to be developed. Herein, we report for the first time the use of droplets Cas13a to detect the circWDR37 as a biomarker of CRC. The arraystar circRNA microarray assays, functional experiments in vitro and in vivo, and qPCR were performed to discover and validate that circWDR37 is a biomarker for early screening and prognosis evaluation of CRC. A new technology named µDCR, which accurately detects circWDR37, has been developed by combining microfluidic droplets with CRISPR/Cas13a and recombinase polymerase amplification (RPA). Meanwhile, the role of crowding agent in improving the performance of Cas13a was uncovered. The 4% polyethylene glycol 8000 and 3% dextran‐10 significantly improved the response speed and sensitivity of one‐pot Cas13a‐RPA reaction. The detection limit of circWDR37 by µDCR was found to be 10 copies/mL, which is higher than that of qPCR. The clinical sample findings demonstrated that circWDR37 detection can be utilized to effectively screen for CRC at an early stage and enable accurate assessment of prognosis. CircWDR37 is confirmed as a groundbreaking biomarker for both diagnosis and prognosis evaluation in CRC patients. Furthermore, our innovative µDCR method for detecting circWDR37 demonstrates impressive attributes such as streamlined operation, rapidity, and high‐throughput, making it an optimal technology platform for the noninvasive screening of CRC. |
| format | Article |
| id | doaj-art-e9c4a3f3ae8948588f785735c0e42ded |
| institution | Kabale University |
| issn | 2692-4560 |
| language | English |
| publishDate | 2025-01-01 |
| publisher | Wiley |
| record_format | Article |
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| spelling | doaj-art-e9c4a3f3ae8948588f785735c0e42ded2025-01-21T08:57:07ZengWileyAggregate2692-45602025-01-0161n/an/a10.1002/agt2.663Droplets Cas13a‐RPA measurement delineates potential role for plasma circWDR37 in colorectal cancerJingsong Xu0Li Cao1Shuang Yang2Ying Jian3Yu Liu4Zhen Shen5Qian Liu6Xiang Chen7Min Li8Shun Li9Xiaolei Zuo10Min Li11Hua Wang12Department of Laboratory Medicine, Renji Hospital, School of Medicine Shanghai Jiao Tong University Shanghai ChinaDepartment of Pathology, The Fifth People's Hospital of Shanghai Fudan University Shanghai ChinaDepartment of Laboratory Medicine, Renji Hospital, School of Medicine Shanghai Jiao Tong University Shanghai ChinaDepartment of Laboratory Medicine, Renji Hospital, School of Medicine Shanghai Jiao Tong University Shanghai ChinaDepartment of Laboratory Medicine, Renji Hospital, School of Medicine Shanghai Jiao Tong University Shanghai ChinaDepartment of Laboratory Medicine, Renji Hospital, School of Medicine Shanghai Jiao Tong University Shanghai ChinaDepartment of Laboratory Medicine, Renji Hospital, School of Medicine Shanghai Jiao Tong University Shanghai ChinaNational Key Laboratory of Science and Technology on Micro/Nano Fabrication Department of Micro/Nano Electronics Shanghai Jiao Tong University Shanghai ChinaInstitute of Molecular Medicine, Renji Hospital, School of Medicine Shanghai Jiao Tong University Shanghai ChinaShanghai Public Health Clinical Center Fudan University Shanghai ChinaInstitute of Molecular Medicine, Renji Hospital, School of Medicine Shanghai Jiao Tong University Shanghai ChinaDepartment of Laboratory Medicine, Renji Hospital, School of Medicine Shanghai Jiao Tong University Shanghai ChinaDepartment of Laboratory Medicine, Renji Hospital, School of Medicine Shanghai Jiao Tong University Shanghai ChinaAbstract Colorectal cancer (CRC) is one of the most prevalent forms of cancer. CircRNAs have emerged as promising biomarkers for cancer diagnosis and prognosis evaluation. However, novel circRNAs as potential biomarkers for CRC still need further exploration and validation, and precise detection methods are yet to be developed. Herein, we report for the first time the use of droplets Cas13a to detect the circWDR37 as a biomarker of CRC. The arraystar circRNA microarray assays, functional experiments in vitro and in vivo, and qPCR were performed to discover and validate that circWDR37 is a biomarker for early screening and prognosis evaluation of CRC. A new technology named µDCR, which accurately detects circWDR37, has been developed by combining microfluidic droplets with CRISPR/Cas13a and recombinase polymerase amplification (RPA). Meanwhile, the role of crowding agent in improving the performance of Cas13a was uncovered. The 4% polyethylene glycol 8000 and 3% dextran‐10 significantly improved the response speed and sensitivity of one‐pot Cas13a‐RPA reaction. The detection limit of circWDR37 by µDCR was found to be 10 copies/mL, which is higher than that of qPCR. The clinical sample findings demonstrated that circWDR37 detection can be utilized to effectively screen for CRC at an early stage and enable accurate assessment of prognosis. CircWDR37 is confirmed as a groundbreaking biomarker for both diagnosis and prognosis evaluation in CRC patients. Furthermore, our innovative µDCR method for detecting circWDR37 demonstrates impressive attributes such as streamlined operation, rapidity, and high‐throughput, making it an optimal technology platform for the noninvasive screening of CRC.https://doi.org/10.1002/agt2.663circRNAcolorectal cancerCRISPR/Cas13acrowding agentdroplet |
| spellingShingle | Jingsong Xu Li Cao Shuang Yang Ying Jian Yu Liu Zhen Shen Qian Liu Xiang Chen Min Li Shun Li Xiaolei Zuo Min Li Hua Wang Droplets Cas13a‐RPA measurement delineates potential role for plasma circWDR37 in colorectal cancer Aggregate circRNA colorectal cancer CRISPR/Cas13a crowding agent droplet |
| title | Droplets Cas13a‐RPA measurement delineates potential role for plasma circWDR37 in colorectal cancer |
| title_full | Droplets Cas13a‐RPA measurement delineates potential role for plasma circWDR37 in colorectal cancer |
| title_fullStr | Droplets Cas13a‐RPA measurement delineates potential role for plasma circWDR37 in colorectal cancer |
| title_full_unstemmed | Droplets Cas13a‐RPA measurement delineates potential role for plasma circWDR37 in colorectal cancer |
| title_short | Droplets Cas13a‐RPA measurement delineates potential role for plasma circWDR37 in colorectal cancer |
| title_sort | droplets cas13a rpa measurement delineates potential role for plasma circwdr37 in colorectal cancer |
| topic | circRNA colorectal cancer CRISPR/Cas13a crowding agent droplet |
| url | https://doi.org/10.1002/agt2.663 |
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