Engineering Gene-Specific DNAzymes for Accessible and Multiplexed Nucleic Acid Testing Article Swipe
Lu Gao
,
Ke Yi
,
Yun Tan
,
Guo Chen
,
Danxi Zheng
,
Chenlan Shen
,
Feng Li
·
YOU?
·
· 2024
· Open Access
·
· DOI: https://doi.org/10.1021/jacsau.4c00232
YOU?
·
· 2024
· Open Access
·
· DOI: https://doi.org/10.1021/jacsau.4c00232
The accurate and timely detection of disease biomarkers at the point-of-care is essential to ensuring effective treatment and epidemiological surveillance. Here, we report the selection and engineering of RNA-cleaving DNAzymes that respond to specific genetic markers and amplify detection signals. Because the target-specific activation of gene-specific DNAzymes (gDz) is like the trans-cleavage activity of clustered regularly interspaced short palindromic repeats (CRISPR) CRISPR-associated (Cas) machinery, we further developed a CRISPR-like assay using RNA-cleaving DNAzyme coupled with isothermal sequence and signal amplification (CLARISSA) for nucleic acid detection in clinical samples. Building on the high sequence specificity and orthogonality of gDzs, CLARISSA is highly versatile and expandable for multiplex testing. Upon integration with an isothermal recombinase polymerase amplification, CLARISSA enabled the detection of human papillomavirus (HPV) 16 in 189 cervical samples collected from cervical cancer screening participants (n = 189) with 100% sensitivity and 97.4% specificity, respectively. A multiplexed CLARISSA further allowed the simultaneous analyses of HPV16 and HPV18 in 46 cervical samples, which returned clinical sensitivity of 96.3% for HPV16 and 83.3% for HPV18, respectively. No false positives were found throughout our tests. Besides the fluorescence readout using fluorogenic reporter probes, CLARISSA is also demonstrated to be fully compatible with a visual lateral flow readout. Because of the high sensitivity, accessibility, and multiplexity, we believe CLARISSA is an ideal CRISPR-Dx alternative for clinical diagnosis in field-based and point-of-care applications.
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- Type
- article
- Language
- en
- Landing Page
- https://doi.org/10.1021/jacsau.4c00232
- https://pubs.acs.org/doi/pdf/10.1021/jacsau.4c00232
- OA Status
- gold
- Cited By
- 13
- References
- 44
- Related Works
- 10
- OpenAlex ID
- https://openalex.org/W4394611206
All OpenAlex metadata
Raw OpenAlex JSON
- OpenAlex ID
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https://openalex.org/W4394611206Canonical identifier for this work in OpenAlex
- DOI
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https://doi.org/10.1021/jacsau.4c00232Digital Object Identifier
- Title
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Engineering Gene-Specific DNAzymes for Accessible and Multiplexed Nucleic Acid TestingWork title
- Type
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articleOpenAlex work type
- Language
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enPrimary language
- Publication year
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2024Year of publication
- Publication date
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2024-04-09Full publication date if available
- Authors
-
Lu Gao, Ke Yi, Yun Tan, Guo Chen, Danxi Zheng, Chenlan Shen, Feng LiList of authors in order
- Landing page
-
https://doi.org/10.1021/jacsau.4c00232Publisher landing page
- PDF URL
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https://pubs.acs.org/doi/pdf/10.1021/jacsau.4c00232Direct link to full text PDF
- Open access
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YesWhether a free full text is available
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goldOpen access status per OpenAlex
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https://pubs.acs.org/doi/pdf/10.1021/jacsau.4c00232Direct OA link when available
- Concepts
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Deoxyribozyme, Nucleic acid, Computational biology, Multiplexing, Computer science, DNA, Biology, Genetics, TelecommunicationsTop concepts (fields/topics) attached by OpenAlex
- Cited by
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13Total citation count in OpenAlex
- Citations by year (recent)
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2025: 11, 2024: 2Per-year citation counts (last 5 years)
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44Number of works referenced by this work
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10Other works algorithmically related by OpenAlex
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