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Published on: December 23, 2022
Detection of MicroRNA-155 based on lambda exonuclease selective digestion and CRISPR/cas12a-assisted amplification
Haotian Zhang1, Jun Xu1, Shiwen Liu2
1Key Lab of Fluorine and Silicon for Energy Materials and Chemistry of Ministry of Education, College of Chemistry and Chemical Engineering, Jiangxi Normal University, Nanchang, 330022, PR China.
Abstract:
In numerous malignancies, miRNA-155 is overexpressed and has oncogenic activity because it is one of the most efficient microRNAs for inhibiting apoptosis in human cancer cells. As a result, the highest sensitive detection of the miRNA-155 gene is a technological instrument that can enable early cancer screening. In this study, a miRNA-155 biosensor was created to create a hairpin probe that can bind to the miRNA-155 gene using lambda nucleic acid exonuclease, which can cut the 5' phosphorylated double strand, and by the DNA probe is recognized by the Cas12a enzyme, which then activates Cas12a to catalyze trans-cutting produces strong fluorescence. Research finding, the target concentration's logarithm and corresponding fluorescence intensity have a strong linear connection, and the limit of detection (LOD) of the sensing system was determined to be 8.3 pM. In addition, the biosensor displayed exceptional specificity, low false-positive signal, and high sensitivity in detecting the miRNA-155 gene in serum samples. This study's creation of a biosensor that has high sensitivity, good selectivity, and is simple to operate provides promising opportunities for research into biosensor design and early cancer detection.
Insights
A novel biosensor effectively detects the miRNA-155 gene, crucial in many cancers. This highly sensitive and specific tool shows promise for early cancer screening using fluorescence detection.
Area of Science:
- Biotechnology
- Molecular Biology
- Cancer Research
Background:
- MicroRNA-155 (miRNA-155) is overexpressed in many cancers, promoting oncogenesis by inhibiting apoptosis.
- Sensitive detection of miRNA-155 is vital for early cancer screening and diagnosis.
Purpose of the Study:
- To develop a highly sensitive and specific biosensor for detecting the miRNA-155 gene.
- To establish a fluorescence-based detection method for miRNA-155.
Main Methods:
- A hairpin probe was designed to bind the miRNA-155 gene.
- Lambda nucleic acid exonuclease was used to cleave the 5' phosphorylated double strand.
- Cas12a enzyme was activated by a DNA probe to catalyze trans-cutting, generating fluorescence.
Main Results:
- A strong linear correlation was observed between the logarithm of target concentration and fluorescence intensity.
- The limit of detection (LOD) for the miRNA-155 gene was determined to be 8.3 pM.
- The biosensor demonstrated high specificity, low false-positive rates, and excellent sensitivity in serum samples.
Conclusions:
- The developed biosensor offers a sensitive, selective, and user-friendly method for miRNA-155 detection.
- This biosensor holds significant potential for advancing early cancer detection strategies.
