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An In Vitro Protocol for Evaluating MicroRNA Levels, Functions, and Associated Target Genes in Tumor Cells
Published on: May 21, 2019
Current updates regarding biogenesis, functions and dysregulation of microRNAs in cancer: Innovative approaches for
Abdulaziz A Aloliqi1, Abdullah M Alnuqaydan1, Aqel Albutti1
1Department of Basic Health Sciences, College of Applied Medical Sciences, Qassim University, Buraydah, Al‑Qassim 51452, Saudi Arabia.
Abstract:
MicroRNAs (miRNAs) are short non‑coding RNAs, which perform a key role in cellular differentiation and development. Most human diseases, particularly cancer, are linked to miRNA functional dysregulation implicated in the expression of tumor‑suppressive or oncogenic targets. Cancer hallmarks such as continued proliferative signaling, dodging growth suppressors, invasion and metastasis, triggering angiogenesis, and avoiding cell death have all been demonstrated to be affected by dysregulated miRNAs. Thus, for the treatment of different cancer types, the detection and quantification of this type of RNA is significant. The classical and current methods of RNA detection, including northern blotting, reverse transcription‑quantitative PCR, rolling circle amplification and next‑generation sequencing, may be effective but differ in efficiency and accuracy. Furthermore, these approaches are expensive, and require special instrumentation and expertise. Thus, researchers are constantly looking for more innovative approaches for miRNA detection, which can be advantageous in all aspects. In this regard, an RNA manipulation tool known as the CRISPR and CRISPR‑associated sequence 13 (CRISPR/Cas13) system has been found to be more advantageous in miRNA detection. The Cas13‑based miRNA detection approach is cost effective and requires no special instrumentation or expertise. However, more research and validation are required to confirm the growing body of CRISPR/Cas13‑based research that has identified miRNAs as possible cancer biomarkers for diagnosis and prognosis, and as targets for treatment. In the present review, current updates regarding miRNA biogenesis, structural and functional aspects, and miRNA dysregulation during cancer are described. In addition, novel approaches using the CRISPR/Cas13 system as a next‑generation tool for miRNA detection are discussed. Furthermore, challenges and prospects of CRISPR/Cas13‑based miRNA detection approaches are described.
Insights
MicroRNAs (miRNAs) are crucial in cell development and cancer. The CRISPR/Cas13 system offers a cost-effective, innovative method for detecting these microRNAs, aiding cancer diagnosis and treatment.
Area of Science:
- Molecular Biology
- Genetics
- Biotechnology
Background:
- MicroRNAs (miRNAs) are key regulators of cellular processes, and their dysregulation is linked to cancer development.
- Cancer hallmarks are significantly influenced by altered miRNA expression, making miRNA detection vital for oncology.
- Current miRNA detection methods face limitations in cost, efficiency, and accessibility.
Purpose of the Study:
- To review current knowledge on miRNA biogenesis, function, and dysregulation in cancer.
- To explore novel CRISPR/Cas13-based systems for miRNA detection.
- To discuss the challenges and future prospects of CRISPR/Cas13 for miRNA analysis.
Main Methods:
- Review of existing literature on miRNA biology and detection techniques.
- Analysis of the application and advantages of CRISPR/Cas13 systems in miRNA detection.
- Discussion of comparative performance against traditional methods like qPCR and NGS.
Main Results:
- CRISPR/Cas13 systems present a promising, cost-effective, and accessible alternative for miRNA detection.
- CRISPR/Cas13-based approaches show potential for identifying miRNAs as cancer biomarkers.
- The review consolidates information on miRNA dysregulation and highlights CRISPR/Cas13's potential in cancer diagnostics.
Conclusions:
- CRISPR/Cas13 technology offers significant advantages for miRNA detection, particularly in cancer research.
- Further research and validation are needed to fully establish CRISPR/Cas13-based miRNA detection for clinical applications.
- This technology holds promise for advancing cancer diagnosis, prognosis, and therapeutic strategies.
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