The DOI matches its Crossref record; 44 of the 44 references it cites were found in a registry or an index.
Clustered Regularly Interspaced Short Palindromic Repeats/Cas9 Triggered Isothermal Amplification for Site-Specific Nucleic Acid Detection
Abstract
A novel CRISPR/Cas9 triggered isothermal exponential amplification reaction (CAS-EXPAR) strategy based on CRISPR/Cas9 cleavage and nicking endonuclease (NEase) mediated nucleic acids amplification was developed for rapid and site-specific nucleic acid detection. CAS-EXPAR was primed by the target DNA fragment produced by cleavage of CRISPR/Cas9, and the amplification reaction performed cyclically to generate a large number of DNA replicates which were detected using a real-time fluorescence monitoring method. This strategy that combines the advantages of CRISPR/Cas9 and exponential amplification showed high specificity as well as rapid amplification kinetics. Unlike conventional nucleic acids amplification reactions, CAS-EXPAR does not require exogenous primers, which often cause target-independent amplification. Instead, primers are first generated by Cas9/sgRNA directed site-specific cleavage of target and accumulated during the reaction. It was demonstrated this strategy gave a detection limit of 0.82 amol and showed excellent specificity in discriminating single-base mismatch. Moreover, the applicability of this method to detect DNA methylation and L. monocytogenes total RNA was also verified. Therefore, CAS-EXPAR may provide a new paradigm for efficient nucleic acid amplification and hold the potential for molecular diagnostic applications.
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Cited by
Kaminski, M. M., Abudayyeh, O. O., Gootenberg, J. S., Zhang, F., & Collins, J. J. (2021) · Nature Biomedical Engineering · doi:10.1038/s41551-021-00760-7 · Cited by 1,259
Cytokines: From Clinical Significance to Quantification
Liu, C., Chu, D., Kalantar‐Zadeh, K., George, J., Young, H. A., & Liu, G. (2021) · Advanced Science · doi:10.1002/advs.202004433 · Cited by 940
CRISPR/Cas Systems towards Next-Generation Biosensing
Li, Y., Li, S., Wang, J., & Liu, G. (2019) · Trends in Biotechnology · doi:10.1016/j.tibtech.2018.12.005 · Cited by 917
Ultrasensitive and Highly Specific Lateral Flow Assays for Point-of-Care Diagnosis
Liu, Y., Zhan, L., Qin, Z., Sackrison, J., & Bischof, J. C. (2021) · ACS Nano · doi:10.1021/acsnano.0c10035 · Cited by 626
Feng, W., Newbigging, A. M., Tao, J., Cao, Y., Peng, H., Le, C., Wu, J., Pang, B., Li, J., Tyrrell, D. L., et al. (2021) · Chemical Science · doi:10.1039/d0sc06973f · Cited by 312
Metadata
- Authors
- Huang, M., Zhou, X., Wang, H., & Xing, D.
- Venue
- Analytical Chemistry
- Publisher
- American Chemical Society (ACS)
- Published
- 2018-01-05
- Volume
- 90
- Issue
- 3
- Pages
- 2193-2200
- ISSN
- 0003-2700 (ISSN-L)
- 1520-6882
- OpenAlex
- W2777258006
- Cited by
- 284
- References
- 44
- FWCI
- 4.76
- Type
- journal article
- Language
- en
- Open access
- closed
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- Huang, M., Zhou, X., Wang, H., & Xing, D. (2018). Clustered Regularly Interspaced Short Palindromic Repeats/Cas9 Triggered Isothermal Amplification for Site-Specific Nucleic Acid Detection. Analytical Chemistry. https://doi.org/10.1021/acs.analchem.7b04542
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