Development of an inducible Cas9 nickase and PAM-free Cas12a platform for bacterial diagnostics
文献类型：期刊论文
| 作者 | Hu, Yuanzhao1; Qiao, Yuefeng1; Li, Xiu-Qing2,5; Xiang, Zhenbo4 ; Wan, Yi1; Wang, Peng3; Yang, Zhiqing1,4
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| 刊名 | TALANTA
 |
| 出版日期 | 2023-12-01 |
| 卷号 | 265页码:6 |
| ISSN号 | 0039-9140 |
| 关键词 | Bacteria diagnostics CRISPR-Cas9n CRISPR-Cas12a Strand displacement |
| DOI | 10.1016/j.talanta.2023.124931 |
| 通讯作者 | Wang, Peng(wangpeng@qdio.ac.cn) ; Yang, Zhiqing(yangzhiqing@hainanu.edu.cn) |
| 英文摘要 | Rapid, efficient, specific and sensitive diagnostic techniques are critical for selecting appropriate treatments for drug-resistant bacterial infections. To address this challenge, we have developed a novel diagnostic method, called the Dual-Cas Tandem Diagnostic Platform (DTDP), which combines the use of Cas9 nickase (Cas9n) and Cas12a. DTDP works by utilizing the Cas9n-sgRNA complex to create a nick in the target strand's doublestranded DNA (dsDNA). This prompts DNA polymerase to displace the single-stranded DNA (ssDNA) and leads to cycles of DNA replication through nicking, displacement, and extension. The ssDNA is then detected by the Cas12a-crRNA complex (which is PAM-free), activating trans-cleavage and generating a fluorescent signal from the fluorescent reporter. DTDP exhibits a high sensitivity (1 CFU/mL or 100 ag/& mu;L), high specificity (specifically to MRSA in nine pathogenic species), and excellent accuracy (100%). The dual RNA recognition process in our method improves diagnostic specificity by decreasing the limitations of Cas12a in detecting dsDNA protospacer adjacent motifs (PAMs) and leverages multiple advantages of multi-Cas enzymes in diagnostics. This novel approach to pathogenic microorganism detection has also great potential for clinical diagnosis. |
| WOS关键词 | INFECTIOUS-DISEASES ; LIMITATIONS ; ADVANTAGES |
| 资助项目 | National Key Ramp;D Program of China[2022YFD2401304] ; Hainan Province Science and Technology Special Fund[ZDYF2023XDNY074] ; Research Foundation of Hainan University ; Hainan Provincial Nat-ural Science Foundation of China[322QN228] ; Hainan Provincial Nat-ural Science Foundation of China[KYQD (ZR) 1997] ; [322MS032] |
| WOS研究方向 | Chemistry |
| 语种 | 英语 |
| 出版者 | ELSEVIER |
| WOS记录号 | WOS:001048130900001 |
| 源URL | [http://ir.qdio.ac.cn/handle/337002/182807]  |
| 专题 | 海洋研究所_海洋腐蚀与防护研究发展中心 |
| 通讯作者 | Wang, Peng; Yang, Zhiqing |
| 作者单位 | 1.Hainan Univ, Marine Coll, State Key Lab Marine Resource Utilizat South China, Haikou 570228, Peoples R China 2.Nutra Hlth Prod & Technol Inc, Fredericton, NB E3B 6J5, Canada 3.Chinese Acad Sci, Inst Oceanol, CAS Key Lab Marine Environm Corros & Biofouling, Qingdao 266071, Peoples R China 4.Rizhao Sci & Technol Innovat Serv Ctr, 369 Jining Rd, Rizhao, Shandong, Peoples R China 5.Agr & Agrifood Canada, Fredericton, NB E3B 4Z7, Canada |
| 推荐引用方式 GB/T 7714 | Hu, Yuanzhao,Qiao, Yuefeng,Li, Xiu-Qing,et al. Development of an inducible Cas9 nickase and PAM-free Cas12a platform for bacterial diagnostics[J]. TALANTA,2023,265:6. |
| APA | Hu, Yuanzhao.,Qiao, Yuefeng.,Li, Xiu-Qing.,Xiang, Zhenbo.,Wan, Yi.,...&Yang, Zhiqing.(2023).Development of an inducible Cas9 nickase and PAM-free Cas12a platform for bacterial diagnostics.TALANTA,265,6. |
| MLA | Hu, Yuanzhao,et al."Development of an inducible Cas9 nickase and PAM-free Cas12a platform for bacterial diagnostics".TALANTA 265(2023):6. |
入库方式： OAI收割
来源：海洋研究所
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