期刊
SENSORS AND ACTUATORS B-CHEMICAL
卷 355, 期 -, 页码 -出版社
ELSEVIER SCIENCE SA
DOI: 10.1016/j.snb.2021.131112
关键词
CRISPR; Cas12a; Hand-in-hand assembly; Leukemia; Cascade signal amplification; Fluorescence sensor
资金
- National Natural Science Foundation of China (NSFC) [22164011, 21565015, 21663014]
- Foundation of Jiangxi Educational Committee [GJJ150363]
- State Key Laboratory of Chemical Biosensing Chemometrics
In this paper, a novel CRISPR/Cas12a fluorescent biosensor for leukemia diagnosis is reported. The biosensor shows a good linear relationship and low detection limit, and maintains good detection performance in actual sample analysis. Compared to traditional CRISPR/Cas detection systems, this biosensor is more advanced and can be used in multiple fields.
CRISPR/Cas technology has attracted increasing attention for use in accurate and sensitive nucleic acid detection, and has innovated the field of molecular diagnostics. Because it is programmability and shows high fidelity and effective signal amplification capabilities. Specifically, Cas12a has been evaluated through in-depth research because it triggers collateral trans-cleavage activity for the target. Here, we report a novel CRISPR/Cas12a fluorescent biosensor by in situ hand-in-hand DNA tile assembly for leukemia diagnosis, and it allows an active isothermal amplification for multi-channel collection of CRISPR/cas12a transducers for achieving three-stage signal amplification. When the target gene concentration was in a range from 1 fM to 50 nM, the fluorescence intensity exhibited an excellent linear relationship with the logarithm of the target gene concentration, and the detection limit reached 1 fM. Furthermore, good detection performance can still be maintained in actual sample analysis. The CRISPR/Cas biosensor for leukemia diagnosis is more advanced than the traditional CRISPR/Cas detection systems, which can be used in cancer gene analyses, kinship judgment, environmental monitoring, and so on.
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