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| Main Authors: | , , , , , , , , , , , , , , |
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| Format: | Artículo científico |
| Language: | en |
| Published: |
Analytical methods : advancing methods and applications
2025
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| Subjects: | |
| Online Access: | https://pubmed.ncbi.nlm.nih.gov/40704969/ |
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| _version_ | 1868266174409605123 |
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| author | Li, Jiaqi Lau, Cia-Hin Wang, Jianchao Wu, Weidong Huang, Zhihao Chen, Xiaoqing Li, Jiahui Huang, Yumei Wang, Tao Li, Yulin Zhao, Zihan Xu, Meijing Chen, Gang Tong, Sheng Zhu, Haibao |
| author_facet | Li, Jiaqi Lau, Cia-Hin Wang, Jianchao Wu, Weidong Huang, Zhihao Chen, Xiaoqing Li, Jiahui Huang, Yumei Wang, Tao Li, Yulin Zhao, Zihan Xu, Meijing Chen, Gang Tong, Sheng Zhu, Haibao Li, Jiaqi Lau, Cia-Hin Wang, Jianchao Wu, Weidong Huang, Zhihao Chen, Xiaoqing Li, Jiahui Huang, Yumei Wang, Tao Li, Yulin Zhao, Zihan Xu, Meijing Chen, Gang Tong, Sheng Zhu, Haibao |
| collection | PubMed - marine biology |
| contents | Rapid, multiplex, one-pot CRISPR/Dx system for detecting cancer fusion genes. Li, Jiaqi Lau, Cia-Hin Wang, Jianchao Wu, Weidong Huang, Zhihao Chen, Xiaoqing Li, Jiahui Huang, Yumei Wang, Tao Li, Yulin Zhao, Zihan Xu, Meijing Chen, Gang Tong, Sheng Zhu, Haibao Humans CRISPR-Cas Systems Lung Neoplasms Proto-Oncogene Proteins Oncogene Proteins, Fusion Protein-Tyrosine Kinases Gene Fusion Targeted therapies directed at fusion genes have proven remarkably effective against cancers. Therefore, the rapid and reliable identification of cancer fusion genes can guide subsequent therapeutic treatment and predict prognosis. By integrating the RT-RPA and CRISPR/Cas12a approaches, we developed a one-pot CRISPR/Dx system for the rapid and multiplex detection of cancer fusion genes. A tube with unique assemblies was created using 3D printing technology to realize this application. As proof of principle, we demonstrated the feasibility of the one-pot CRISPR/Dx system in detecting lung cancer by targeting fusions. The performance of the one-pot CRISPR/Dx detection system was comparable to a two-tube-based testing platform. When tested with synthetic RNA fusions, both approaches efficiently detected all 14 fusions with an LOD in the range of 5-10 copies per μL, without generating a background signal, even in the presence of a large excess of wild-type RNA. The total reaction time for both approaches was 30 minutes. Notably, the one-pot CRISPR/Dx detection system minimized the operation steps and aerosol contamination without compromising detection sensitivity and specificity. Furthermore, its diagnostic power was validated using clinical samples. Thus, we successfully developed a rapid, multiplex, one-pot CRISPR/Dx detection system for detecting 14 clinically relevant fusions with high sensitivity and specificity. It is also cost-effective and simple to operate, thereby realizing the ultimate goal of establishing CRISPR/Dx as the paragon of cancer diagnostics for home self-testing and point-of-care testing. |
| format | Artículo científico |
| id | pubmed_40704969 |
| institution | PubMed |
| language | en |
| publishDate | 2025 |
| publisher | Analytical methods : advancing methods and applications |
| record_format | pubmed |
| spellingShingle | Rapid, multiplex, one-pot CRISPR/Dx system for detecting cancer fusion genes. Li, Jiaqi Lau, Cia-Hin Wang, Jianchao Wu, Weidong Huang, Zhihao Chen, Xiaoqing Li, Jiahui Huang, Yumei Wang, Tao Li, Yulin Zhao, Zihan Xu, Meijing Chen, Gang Tong, Sheng Zhu, Haibao Humans CRISPR-Cas Systems Lung Neoplasms Proto-Oncogene Proteins Oncogene Proteins, Fusion Protein-Tyrosine Kinases Gene Fusion Rapid, multiplex, one-pot CRISPR/Dx system for detecting cancer fusion genes. Li, Jiaqi Lau, Cia-Hin Wang, Jianchao Wu, Weidong Huang, Zhihao Chen, Xiaoqing Li, Jiahui Huang, Yumei Wang, Tao Li, Yulin Zhao, Zihan Xu, Meijing Chen, Gang Tong, Sheng Zhu, Haibao Humans CRISPR-Cas Systems Lung Neoplasms Proto-Oncogene Proteins Oncogene Proteins, Fusion Protein-Tyrosine Kinases Gene Fusion Targeted therapies directed at fusion genes have proven remarkably effective against cancers. Therefore, the rapid and reliable identification of cancer fusion genes can guide subsequent therapeutic treatment and predict prognosis. By integrating the RT-RPA and CRISPR/Cas12a approaches, we developed a one-pot CRISPR/Dx system for the rapid and multiplex detection of cancer fusion genes. A tube with unique assemblies was created using 3D printing technology to realize this application. As proof of principle, we demonstrated the feasibility of the one-pot CRISPR/Dx system in detecting lung cancer by targeting fusions. The performance of the one-pot CRISPR/Dx detection system was comparable to a two-tube-based testing platform. When tested with synthetic RNA fusions, both approaches efficiently detected all 14 fusions with an LOD in the range of 5-10 copies per μL, without generating a background signal, even in the presence of a large excess of wild-type RNA. The total reaction time for both approaches was 30 minutes. Notably, the one-pot CRISPR/Dx detection system minimized the operation steps and aerosol contamination without compromising detection sensitivity and specificity. Furthermore, its diagnostic power was validated using clinical samples. Thus, we successfully developed a rapid, multiplex, one-pot CRISPR/Dx detection system for detecting 14 clinically relevant fusions with high sensitivity and specificity. It is also cost-effective and simple to operate, thereby realizing the ultimate goal of establishing CRISPR/Dx as the paragon of cancer diagnostics for home self-testing and point-of-care testing. |
| title | Rapid, multiplex, one-pot CRISPR/Dx system for detecting cancer fusion genes. |
| topic | Humans CRISPR-Cas Systems Lung Neoplasms Proto-Oncogene Proteins Oncogene Proteins, Fusion Protein-Tyrosine Kinases Gene Fusion |
| url | https://pubmed.ncbi.nlm.nih.gov/40704969/ |