Advances in CRISPR based biosensing strategies for cancer diagnosis
image: (a) Utilizing HCR and CRISPR-Cas12a double amplification technique, a highly sensitive apta-HCR-CRISPR method was devised for the detection of TEV protein. (b) A novel CRISPR-Cas12a/Cas13a approach, activated by DNAzyme walkers, is suggested for the concurrent identification of exosomal proteins such as SAA1 and clotting FV. (c) A fluorescence aptasensor was created to detect NPC-derived exosomes by utilizing a combination of MNPs, TdT, and CRISPR/Cas12a. (d) An innovative biosensor, incorporating MNPs, CSDR, and CRISPR/Cas12a, was created to identify the exosome miR-21 in lung cancer. Cas12a, CRISPR-associated protein 12a; Cas13a, CRISPR-associated protein 13a; CD, Cluster of Differentiation; CRISPR, Clustered Regularly Interspaced Short Palindromic Repeats; crRNA, CRISPR RNA; DL, Deep Learning; EVs, Extracellular Vesicles; FQ, Fluoroquinolone;PD-L1, Programmed Death-Ligand 1. (a) Copyright 2020, Theranostics. (b) Copyright(2023), with permission from Elsevier; (c) Reproduced with permission from Springer Nature.(d) Copyright 2022, Multidisciplinary Digital Publishing Institute.
Credit: Shan-Wen Hu, Qi-Ling Yang
With the rapid development of gene editing technology, its applications in tumor diagnosis have expanded significantly. The CRISPR/Cas system, a crucial gene editing tool, enhances the early detection and precision diagnosis of tumors, facilitating high-throughput and high-sensitivity detection. This review focuses on the various applications of the CRISPR/Cas system in tumor diagnosis, highlighting advancements and future prospects.
Detection of Tumor-derived Exosomes
Exosomes, small extracellular vesicles secreted by tumor cells, play a vital role in tumor communication and metastasis. The CRISPR/Cas system can identify surface proteins or nucleic acids of these exosomes, enabling non-invasive liquid biopsies. By utilizing adaptors that recognize these exosome markers, the CRISPR system is activated, providing a precise method for detecting and monitoring tumors in real-time. This technique holds promise for early diagnosis and treatment monitoring without the need for invasive procedures.
Detection of Circulating Tumor DNA
Circulating tumor DNA (ctDNA) is fragmented DNA released by tumor cells into the bloodstream. The CRISPR/Cas system can detect ctDNA with high sensitivity, enabling the monitoring of tumor dynamics. Various strategies, such as the spherical nucleic acid reporter-based cascade CRISPR/Cas12a amplifier and proximity hybridization-regulated CRISPR/Cas12a-based dual signal amplification, have been developed to enhance the stability and sensitivity of ctDNA detection. These methods achieve detection limits as low as 5.43 fM, facilitating the early detection of tumors and the assessment of treatment efficacy.
Detection of Circulating Tumor Cells
Circulating tumor cells (CTCs) are cancer cells that have shed from the primary tumor into the bloodstream and are responsible for metastasis. The CRISPR/Cas system, combined with aptamers that recognize tumor-specific surface proteins, allows for the detection of CTCs even in trace amounts. Techniques like the Multivalent Duplexed-Aptamer Networks Regulated CRISPR-Cas12a system and FINDER (Fluidly Confined CRISPR-Based DNA Reporter) have shown high efficiency in identifying and quantifying CTCs. These methods achieve detection limits as low as 26 cells/ml, providing valuable insights into tumor progression and metastasis.
Detection of Tumor Biomarkers
Tumor markers are biomolecules produced by tumor cells that can be detected in blood, urine, or tissues. The CRISPR/Cas system enhances the sensitivity and specificity of tumor marker detection through various innovative approaches. For example, platinum nanoparticles-based CRISPR/Cas12a platforms and CRISPR-powered biosensing platforms for alpha-fetoprotein detection have shown remarkable sensitivity, with detection limits as low as 0.01 ng/mL. These advancements enable accurate tumor diagnosis and monitoring, aiding in the personalized treatment of cancer patients.
Detection and Identification of Tumor Microenvironments
The tumor microenvironment (TME) consists of various non-cancerous cells and molecules that surround and interact with tumor cells. Understanding the TME is crucial for developing effective cancer therapies. The CRISPR/Cas system can be utilized to detect and identify components of the TME by activating gene editing enzymes through differential factors present in the TME. This approach allows for the dynamic monitoring of tumors and provides insights into the interactions between tumor cells and their microenvironment, paving the way for targeted therapies.
Discussion
The CRISPR/Cas system has revolutionized tumor diagnosis by providing highly sensitive, specific, and non-invasive detection methods. However, challenges such as off-target effects, delivery efficiency, and regulatory concerns need to be addressed to fully realize its potential. Future research should focus on improving the specificity and efficiency of CRISPR-based detection systems, developing robust delivery mechanisms, and ensuring the clinical applicability of these technologies.
Conclusions
The CRISPR/Cas system offers a transformative approach to tumor diagnosis, enabling early detection, precise monitoring, and personalized treatment strategies. Continued advancements in this field hold great promise for improving cancer diagnosis and patient outcomes. Addressing current challenges will pave the way for the widespread clinical adoption of CRISPR-based diagnostic tools, ultimately enhancing the efficacy of cancer treatment and management.
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https://www.xiahepublishing.com/2835-3315/CSP-2023-00026
The study was recently published in the Cancer Screening and Prevention.
Cancer Screening and Prevention (CSP) publishes high-quality research and review articles related to cancer screening and prevention. It aims to provide a platform for studies that develop innovative and creative strategies and precise models for screening, early detection, and prevention of various cancers. Studies on the integration of precision cancer prevention multiomics where cancer screening, early detection and prevention regimens can precisely reflect the risk of cancer from dissected genomic and environmental parameters are particularly welcome.
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