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Application of CRISPR/Cas12a in miRNA-155 detection: A novel homogeneous electrochemiluminescence biosensor
Yuanxun Gong1, Hongyu Zeng1, Sisi Gao1
1Guangxi Key Laboratory for Preclinical and Translational Research on Bone and Joint Degenerative Diseases, Affiliated Hospital of Youjiang Medical University for Nationalities, Baise, Guangxi, 533000, China.
Background:
MicroRNAs (miRNAs) are important non-coding RNA entities that affect gene expression and function by binding to target mRNAs, leading to degradation of the mRNAs or inhibiting their translation. MiRNAs are widely involved in a variety of biological processes, such as cell differentiation, development, metabolism, and apoptosis. In addition, miRNAs are associated with many diseases, including cancer. However, conventional detection techniques often suffer from shortcomings such as low sensitivity, so we need to develop a rapid and efficient detection strategy for accurate detection of miRNAs.
Results:
We have developed an innovative homogeneous electrochemiluminescence (ECL) biosensor. This biosensor employs CRISPR/Cas12a gene editing technology for accurate and efficient detection of microRNA (miRNA). Compared to conventional technologies, this biosensor employs a unique homogeneous detection format that eliminates laborious probe fixation steps and greatly simplifies the detection process. By using two amplification techniques - isothermal amplification and T7 RNA polymerase amplification - the biosensor improves the sensitivity and specificity of the assay, providing excellent detection performance in the assay. This makes it possible to evaluate miRNA directly from a variety of biological samples such as cell lysates and diluted human serum. Experimental results convincingly demonstrate the extraordinary performance of this biosensor, including its extremely low detection limit of 1.27 aM, high sensitivity, reproducibility and stability.
Significance:
The application of our constructed sensor in distinguishing between cancerous and non-cancerous cell lines highlights its potential for early cancer detection and monitoring. This innovative approach represents a major advancement in the field of miRNA detection, providing a user-friendly, cost-effective, and sensitive solution with broad implications for clinical diagnosis and patient care, especially in point-of-care settings.
Insights
A novel CRISPR/Cas12a-based electrochemiluminescence biosensor offers sensitive and efficient microRNA detection. This method simplifies processes and shows promise for early cancer diagnostics.
Area of Science:
- Biotechnology
- Molecular Biology
- Biosensor Technology
Background:
- MicroRNAs (miRNAs) regulate gene expression and are implicated in various biological processes and diseases like cancer.
- Conventional miRNA detection methods often lack sensitivity and efficiency.
- There is a need for rapid, sensitive, and accurate miRNA detection strategies.
Purpose of the Study:
- To develop an innovative homogeneous electrochemiluminescence (ECL) biosensor for accurate and efficient microRNA (miRNA) detection.
- To overcome the limitations of conventional miRNA detection techniques.
Main Methods:
- Utilized CRISPR/Cas12a gene editing technology for miRNA detection.
- Employed a homogeneous detection format, eliminating laborious probe fixation.
- Incorporated isothermal amplification and T7 RNA polymerase amplification for enhanced sensitivity and specificity.
Main Results:
- Developed a novel homogeneous ECL biosensor for miRNA detection.
- Achieved an extremely low detection limit of 1.27 aM.
- Demonstrated high sensitivity, specificity, reproducibility, and stability in detecting miRNAs from biological samples like cell lysates and human serum.
Conclusions:
- The biosensor shows potential for early cancer detection and monitoring by distinguishing between cancerous and non-cancerous cell lines.
- This approach offers a user-friendly, cost-effective, and sensitive solution for clinical diagnosis.
- The technology has broad implications for patient care, particularly in point-of-care settings.
