DNA Methylation Detection: Fluorescent and Colourimetric Methods Based on Methylation-Associated Restriction Endonucleases

DNA methylation is a key epigenetic alteration that regulates gene expression and plays a vital role in numerous biological processes. Abnormal DNA methylation can lead to various diseases, including cancer. Thus, accurate detection of DNA methylation is crucial for biomedical research. However, conventional methods for methylation detection in clinical settings are slow and tedious. Hence, there is a need for the development of new detection techniques. Fluorescence-based and colorimetric methods have become popular due to their simplicity and rapid response. Furthermore, restriction endonucleases recognise specific DNA sequences and cut the DNA, and are known as the “molecular scissors” of DNA. Methylation-associated restriction endonucleases can selectively cleave DNA based on its methylation status, thereby determining if the target gene is methylated or not. Integration of methylation-associated restriction endonucleases with fluorescence or colorimetric assays offers a fresh approach to DNA methylation detection. This paper reviews traditional detection methods and their drawbacks, emphasizes the use of restriction endonuclease-based fluorescence and colorimetric assays in DNA methylation detection, and discusses existing limitations of these methods along with potential enhancement strategies.