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Active CRISPR-Cas12a on Hydrophilic Metal–Organic Frameworks: A Nanobiocomposite with High Stability and Activity for Nucleic Acid Detection
Analytical Chemistry ( IF 6.7 ) Pub Date : 2023-07-06 , DOI: 10.1021/acs.analchem.3c00400
Zhirun Ji 1 , Bin Zhou 2 , Zhaoyang Shang 1 , Sirui Liu 1 , Xue Li 1 , Xing Zhang 1 , Bingzhi Li 1
Affiliation  

CRISPR-Cas12a is an accurate and responsive biosensing technique, but its limited stability has restricted its widespread applications. To address this, we propose a strategy using metal–organic frameworks (MOFs) to protect Cas12a from harsh environments. After screening multiple candidate MOFs, it was found that hydrophilic MAF-7 is highly compatible with Cas12a, and the as-formed Cas12a-on-MAF-7 (COM) not only retains high enzymatic activity but also possesses excellent tolerance to heat, salt, and organic solvents. Further investigation showed that COM can serve as an analytical component for nucleic acid detection, resulting in an ultrasensitive assay for SARS-CoV-2 RNA detection with a detection limit of 1 copy. This is the first successful attempt to create an active Cas12a nanobiocomposite that functions as a biosensor without the need for shell deconstruction or enzyme release.

中文翻译:

亲水性金属有机框架上的活性 CRISPR-Cas12a:一种具有高稳定性和高活性的核酸检测纳米生物复合材料

CRISPR-Cas12a是一种准确且响应灵敏的生物传感技术,但其有限的稳定性限制了其广泛应用。为了解决这个问题,我们提出了一种使用金属有机框架(MOF)来保护 Cas12a 免受恶劣环境影响的策略。经过筛选多个候选MOF,发现亲水性的MAF-7与Cas12a高度相容,形成的C as12a- on - M AF-7(COM)不仅保留了较高的酶活性,而且对Cas12a具有优异的耐受性。热、盐和有机溶剂。进一步的研究表明,COM 可以作为核酸检测的分析组件,从而实现 SARS-CoV-2 RNA 检测的超灵敏测定,检测限为 1 个拷贝。这是创建活性 Cas12a 纳米生物复合材料的首次成功尝试,该复合材料可用作生物传感器,无需解构外壳或释放酶。
更新日期:2023-07-06
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