Building the Future of Clinical Diagnostics: An Analysis of Potential Benefits and Current Barriers in CRISPR/Cas Diagnostics

Overview

Journal ACS Synth Biol

Publisher American Chemical Society

Date 2025 Jan 29

PMID 39880685

Authors

Jeanne E van Dongen

Loes I Segerink

Affiliations

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Abstract

Advancements in molecular diagnostics, such as polymerase chain reaction and next-generation sequencing, have revolutionized disease management and prognosis. Despite these advancements in molecular diagnostics, the field faces challenges due to high operational costs and the need for sophisticated equipment and highly trained personnel besides having several technical limitations. The emergent field of CRISPR/Cas sensing technology is showing promise as a new paradigm in clinical diagnostics, although widespread clinical adoption remains limited. This perspective paper discusses specific cases where CRISPR/Cas technology can surmount the challenges of existing diagnostic methods by stressing the significant role that CRISPR/Cas technology can play in revolutionizing clinical diagnostics. It underscores the urgency and importance of addressing the technological and regulatory hurdles that must be overcome to harness this technology effectively in clinical laboratories.

References

Ghouneimy A, Mahas A, Marsic T, Aman R, Mahfouz M . CRISPR-Based Diagnostics: Challenges and Potential Solutions toward Point-of-Care Applications. ACS Synth Biol. 2022; 12(1):1-16. PMC: 9872163. DOI: 10.1021/acssynbio.2c00496. View

Choi J, Lim J, Shin M, Paek S, Choi J . CRISPR-Cas12a-Based Nucleic Acid Amplification-Free DNA Biosensor via Au Nanoparticle-Assisted Metal-Enhanced Fluorescence and Colorimetric Analysis. Nano Lett. 2020; 21(1):693-699. DOI: 10.1021/acs.nanolett.0c04303. View

Kellner M, Koob J, Gootenberg J, Abudayyeh O, Zhang F . SHERLOCK: nucleic acid detection with CRISPR nucleases. Nat Protoc. 2019; 14(10):2986-3012. PMC: 6956564. DOI: 10.1038/s41596-019-0210-2. View

Waarts M, Stonestrom A, Park Y, Levine R . Targeting mutations in cancer. J Clin Invest. 2022; 132(8). PMC: 9012285. DOI: 10.1172/JCI154943. View

Park J, Hsieh K, Chen L, Kaushik A, Trick A, Wang T . Digital CRISPR/Cas-Assisted Assay for Rapid and Sensitive Detection of SARS-CoV-2. Adv Sci (Weinh). 2021; 8(5):2003564. PMC: 7927608. DOI: 10.1002/advs.202003564. View

Malci K, Walls L, Rios-Solis L . Rational Design of CRISPR/Cas12a-RPA Based One-Pot COVID-19 Detection with Design of Experiments. ACS Synth Biol. 2022; 11(4):1555-1567. PMC: 9016756. DOI: 10.1021/acssynbio.1c00617. View

Nouri R, Dong M, Politza A, Guan W . Figure of Merit for CRISPR-Based Nucleic Acid-Sensing Systems: Improvement Strategies and Performance Comparison. ACS Sens. 2022; 7(3):900-911. PMC: 9191621. DOI: 10.1021/acssensors.2c00024. View

Hao L, Zhao R, Welch N, Tan E, Zhong Q, Harzallah N . CRISPR-Cas-amplified urinary biomarkers for multiplexed and portable cancer diagnostics. Nat Nanotechnol. 2023; 18(7):798-807. PMC: 10359190. DOI: 10.1038/s41565-023-01372-9. View

Batista A, Pacheco L . Detecting pathogens with Zinc-Finger, TALE and CRISPR- based programmable nucleic acid binding proteins. J Microbiol Methods. 2018; 152:98-104. DOI: 10.1016/j.mimet.2018.07.024. View

10.

Guo L, Sun X, Wang X, Liang C, Jiang H, Gao Q . SARS-CoV-2 detection with CRISPR diagnostics. Cell Discov. 2020; 6:34. PMC: 7235268. DOI: 10.1038/s41421-020-0174-y. View

11.

Shinoda H, Taguchi Y, Nakagawa R, Makino A, Okazaki S, Nakano M . Amplification-free RNA detection with CRISPR-Cas13. Commun Biol. 2021; 4(1):476. PMC: 8055673. DOI: 10.1038/s42003-021-02001-8. View

12.

Bustin S, Benes V, Garson J, Hellemans J, Huggett J, Kubista M . The MIQE guidelines: minimum information for publication of quantitative real-time PCR experiments. Clin Chem. 2009; 55(4):611-22. DOI: 10.1373/clinchem.2008.112797. View

13.

Kleinstiver B, Pattanayak V, Prew M, Tsai S, Nguyen N, Zheng Z . High-fidelity CRISPR-Cas9 nucleases with no detectable genome-wide off-target effects. Nature. 2016; 529(7587):490-5. PMC: 4851738. DOI: 10.1038/nature16526. View

14.

van Dongen J, Berendsen J, Eijkel J, Segerink L . A CRISPR/Cas12a-assisted in vitro diagnostic tool for identification and quantification of single CpG methylation sites. Biosens Bioelectron. 2021; 194:113624. DOI: 10.1016/j.bios.2021.113624. View

15.

Li S, Cheng Q, Wang J, Li X, Zhang Z, Gao S . CRISPR-Cas12a-assisted nucleic acid detection. Cell Discov. 2018; 4:20. PMC: 5913299. DOI: 10.1038/s41421-018-0028-z. View

16.

Chen J, Ma E, Harrington L, Da Costa M, Tian X, Palefsky J . CRISPR-Cas12a target binding unleashes indiscriminate single-stranded DNase activity. Science. 2018; 360(6387):436-439. PMC: 6628903. DOI: 10.1126/science.aar6245. View

17.

Strohkendl I, Saifuddin F, Rybarski J, Finkelstein I, Russell R . Kinetic Basis for DNA Target Specificity of CRISPR-Cas12a. Mol Cell. 2018; 71(5):816-824.e3. PMC: 6679935. DOI: 10.1016/j.molcel.2018.06.043. View

18.

Swarts D, Jinek M . Mechanistic Insights into the cis- and trans-Acting DNase Activities of Cas12a. Mol Cell. 2019; 73(3):589-600.e4. PMC: 6858279. DOI: 10.1016/j.molcel.2018.11.021. View

19.

Xuan J, Yu Y, Qing T, Guo L, Shi L . Next-generation sequencing in the clinic: promises and challenges. Cancer Lett. 2012; 340(2):284-95. PMC: 5739311. DOI: 10.1016/j.canlet.2012.11.025. View

20.

Guo X, Tian T, Deng X, Song Y, Zhou X, Song E . CRISPR/Cas13a assisted amplification of magnetic relaxation switching sensing for accurate detection of miRNA-21 in human serum. Anal Chim Acta. 2022; 1209:339853. DOI: 10.1016/j.aca.2022.339853. View