1. A label-free electrochemical sensor for the detection of two kinds of targets based on CRISPR/Cas12a system.
- Author
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Liu, Bo, Lu, Weishen, Huang, Yibo, Zhang, Xiaoru, and Yuan, Xunyi
- Subjects
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CRISPRS , *ELECTROCHEMICAL sensors , *SINGLE-stranded DNA , *NUCLEIC acids , *EXCHANGE reactions , *DETECTION limit - Abstract
In recent years, electrochemical sensors for nucleic acid testing based on CRISPR/Cas12a technology have attracted widespread interest due to their ease of operation and high detection sensitivity. Here, a label-free electrochemical sensor amplified by primer exchange reaction (PER), hybridization chain reaction (HCR) and CRISPR/Cas12a system was developed for analyzing miRNA- 155 and PCB 77. In the presence of targets, PER was initiated, whose products could be used to trigger the trans-cleavage activity of Cas12a for the single-stranded DNA (ssDNA) on the electrode. Afterwards, with the amplification of HCR on the electrode and electrostatic adsorption of Ru(NH 3) 6 3+, the concentration of the targets could be indicated by the DPV intensity with a low background signal. The detection limits for miRNA- 155 and PCB 77 were estimated to be 67 aM and 6.9 pg/L, respectively. This work provides a versatile method for sensitive detection of different targets using the same crRNA and can be used in various biochemical investigations. • A label-free electrochemical sensor was used for the detection of two targets. • PER, CRISPR/Cas system and HCR were applied for signal amplification. • CRISPR/Cas system can decrease the background signal without the usage of PAN. • The same crRNA can be used to detect different targets. [ABSTRACT FROM AUTHOR]
- Published
- 2024
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