Journal
ACS SENSORS
Volume 5, Issue 6, Pages 1615-1623Publisher
AMER CHEMICAL SOC
DOI: 10.1021/acssensors.0c00081
Keywords
nucleic acid sequence; uracil-DNA glycosylase; strand displacement amplification; fluorescence; CRISPR/Cas12a
Funding
- National Natural Science Foundation of China [81772290, 81271930]
- Graduate Scientific Research and Innovation Foundation of Chongqing, China [CYB19041]
- Chongqing Science and Technology Commission [CSTC2018jcyjAX0062]
- Fundamental Research Funds for the Central Universities [2019CDYGZD007]
- Chongqing Graduate Tutor Team Construction Project
- sharing fund of Chongqing University's large equipment
Ask authors/readers for more resources
Ultrasensitive detection of sequence-specific DNA and uracilDNA glycosylase (UDG) activity shows great practical significance in clinical diagnostic and biomedical studies. Here, a methodology based on a CRISPR/Cas12a system coupled with enhanced strand displacement amplification (ESDA) was innovatively established for sequence-specific DNA or UDG activity detection. Sequence-specific DNA or DNA primers processed by UDG and Endonuclease IV can initiate E-SDA, generating auxiliary DNA chains, which act as activators to unlock the indiscriminate collateral cleavage activities (trans-cleavage) of the CRISPR/Cas12a. Then, the activated CRISPR/Cas12a, which intrinsically possesses the ability of significant signal amplification, can indiscriminately cleave the added cleavage reporters in the system. Thus, the multistep amplification of the method was obtained. Under the selected experimental conditions, the established method can achieve an actual sensitivity of sequence-specific DNA up to 100 aM within 2.5 h or ultralow UDG activity (3.1x10(-5) U/mL) detection within 3.5 h. We believe that the proposed method will have great potential for practical application in ultrasensitive detection of sequence-specific DNA or UDG activity.
Authors
I am an author on this paper
Click your name to claim this paper and add it to your profile.
Reviews
Recommended
No Data Available