Unlocking the potential: Cas13-based sensing system in early detection of micro-RNA biomarker

Autor: Shreya Reddy, Rishita Swami, Jaishiv Ghumde, Athulya Nair, Simran Kushwaha, Shrutam Somkuwar, Devyani Salodkar, Aparna Nair, Deovrat Begde
Jazyk: angličtina
Rok vydání: 2024
Předmět:
Zdroj: Indian Journal of Health Sciences and Biomedical Research KLEU, Vol 17, Iss 3, Pp 234-238 (2024)
Druh dokumentu: article
ISSN: 2542-6214
2542-6222
DOI: 10.4103/kleuhsj.kleuhsj_600_23
Popis: AIM: The aim of this study is to standardize in-vitro CRISPR-Cas13 assay for the detection of salivary microRNAs (miRNAs) as Oral Cancer Biomarker. BACKGROUND: Oral cavity cancer poses a significant global health threat, prevalent in both emerging and industrialized nations, primarily attributed to factors such as tobacco and alcohol use. This study addresses the imperative need for adaptive, effective, and non-invasive strategies in the diagnosis and treatment of oral cancer, emphasising the importance of early detection for improved prognosis. The study introduces a novel approach employing the CRISPR-Cas13 system for the detection of salivary miRNA-145 as an oral cancer biomarker. Material and methods: The study utilized the CRISPR-Cas13 system to detect salivary miRNA-145. Specific guide RNA (gRNA) sequences were designed which guides LwaCas13a and targets miRNA-145, resulting in activation of collateral cleavage activity. Total RNA isolated from salivary samples were used in the assay, and the detection of miRNA-145 was assessed through distinct reduction in band intensity and fluorescence detection using a FAM based reporter RNA. Results: This study has successfully set up CRISPR-Cas13 in-vitro assay for detection of miRNA. The assay demonstrated a distinct reduction in band intensity in test samples compared to controls when total RNA was used. The assay employing reporter RNA illustrates fluorescence detection exclusively in test samples utilizing specific gRNA. Conclusion: This study introduces an innovative method for oral cancer detection, leveraging the CRISPR-Cas13 system and salivary miRNA-145 as a biomarker. The approach presents a non-invasive, cost-effective, and efficient alternative for early cancer diagnosis and monitoring, showcasing its potential impact on advancing diagnostic technologies in the field.
Databáze: Directory of Open Access Journals
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