School of Life Sciences, South China Normal University, Guangzhou, 510631, China.
State Key Laboratory of Oncology in South China, Collaborative Innovation Center for Cancer Medicine, Sun Yat-sen University Cancer Center, Guangzhou, 510060, China.
Angew Chem Int Ed Engl. 2023 Jun 5;62(23):e202300663. doi: 10.1002/anie.202300663. Epub 2023 Apr 27.
The clustered regularly interspaced short palindromic repeats (CRISPR) system is a promising platform for nucleic acid detection. Regulating the CRISPR reaction would be extremely useful to improve the detection efficiency and speed of CRISPR diagnostic applications. Here, we have developed a light-start CRISPR-Cas12a reaction by employing caged CRISPR RNA (crRNA). When combined with recombinase polymerase amplification, a robust photocontrolled one-pot assay is achieved. The photocontrolled one-pot assay is simpler and is 50-fold more sensitive than the conventional assay. This improved detection efficiency also facilitates the development of a faster CRISPR diagnostic method. The detection of clinical samples demonstrated that 10-20 min is sufficient for effective detection, which is much faster than the current gold-standard technique PCR. We expect this advance in CRISPR diagnostics to promote its widespread detection applications in biomedicine, agriculture, and food safety.
成簇规律间隔短回文重复（CRISPR）系统是一种很有前途的核酸检测平台。调控 CRISPR 反应对于提高 CRISPR 诊断应用的检测效率和速度将非常有用。在这里，我们通过使用笼状 CRISPR RNA（crRNA）开发了一种轻启 CRISPR-Cas12a 反应。当与重组酶聚合酶扩增结合时，实现了强大的光控一键式测定。光控一键式测定比传统测定更简单，灵敏度提高了 50 倍。这种改进的检测效率也有助于开发更快的 CRISPR 诊断方法。对临床样本的检测表明，10-20 分钟即可有效检测，比目前的金标准技术 PCR 快得多。我们期望这一 CRISPR 诊断方面的进展将促进其在生物医学、农业和食品安全领域的广泛检测应用。
