期刊
FOOD CHEMISTRY
卷 378, 期 -, 页码 -出版社
ELSEVIER SCI LTD
DOI: 10.1016/j.foodchem.2021.131802
关键词
Lead ion; CRISPR-Cas12a; G-quadruplex; Fluorescence resonance energy transfer
资金
- Science, Technology Commission of Shanghai Municipality [20392001800]
- National Natural Science Foundation of China [31671923]
- National Key Research and Development Plan [2018YFC160440]
In this study, the inhibition of CRISPR-Cas trans-cleavage by Pb2+-induced G-quadruplex was investigated for the detection of Pb2+ in tea beverage and milk. The results showed that Pb2+ competitively combines with G-quadruplex, resulting in its resistance to CRISPR-Cas trans-cleavage. The detection of Pb2+ was achieved through fluorescence resonance energy transfer.
In this work, the inhibition of clustered regularly interspaced short palindromic repeats (CRISPR) - CRISPR associated protein (Cas) trans-cleavage by Pb2+-induced G-quadruplex has been firstly explored to detect Pb2+ in tea beverage and milk. In absence of Pb2+, the Na+-induced G-quadruplex can be cleaved by CRISPR-Cas12a. In contrast, Pb2+ can competitively combine with G-quadruplex, resulting in its conformational changes and resistance to trans-cleavage of CRISPR-Cas12a. Therefore, the fluorescence resonance energy transfer can happen. Pb2+ can be detected in a linear range from 100 nM to 5 mu M with a lowest detection limit of 2.6 nM and a relative standard deviation of 4.32%. In summary, this work not only provides a new method for Pb2+ detection based on its induced G-quadruplex inhibition on CRISPR-Cas12a trans-cleavage, but also broadens the application of CRISPR-Cas system for heavy metal analysis in the field of food safety.
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