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Integrated Micropillar Polydimethylsiloxane Accurate CRISPR Detection System for Viral DNA Sensing

[Image: see text] A fully Integrated Micropillar Polydimethylsiloxane Accurate CRISPR deTection (IMPACT) system is developed for viral DNA detection. This powerful system is patterned with high-aspect-ratio micropillars to enhance reporter probe binding. After surface modification and probe immobili...

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Autores principales: Hass, Kenneth N., Bao, Mengdi, He, Qian, Liu, Li, He, Jiacheng, Park, Myeongkee, Qin, Peiwu, Du, Ke
Formato: Online Artículo Texto
Lenguaje:English
Publicado: American Chemical Society 2020
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7594154/
https://www.ncbi.nlm.nih.gov/pubmed/33134706
http://dx.doi.org/10.1021/acsomega.0c03917
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author Hass, Kenneth N.
Bao, Mengdi
He, Qian
Liu, Li
He, Jiacheng
Park, Myeongkee
Qin, Peiwu
Du, Ke
author_facet Hass, Kenneth N.
Bao, Mengdi
He, Qian
Liu, Li
He, Jiacheng
Park, Myeongkee
Qin, Peiwu
Du, Ke
author_sort Hass, Kenneth N.
collection PubMed
description [Image: see text] A fully Integrated Micropillar Polydimethylsiloxane Accurate CRISPR deTection (IMPACT) system is developed for viral DNA detection. This powerful system is patterned with high-aspect-ratio micropillars to enhance reporter probe binding. After surface modification and probe immobilization, the CRISPR-Cas12a/crRNA complex is injected into the fully enclosed microchannel. With the presence of a double-stranded DNA target, the CRISPR enzyme is activated and denatures the single-stranded DNA reporters from the micropillars. This collateral cleavage releases fluorescence reporters into the assay, and the intensity is linearly proportional to the target DNA concentration ranging from 0.1 to 10 nM. Importantly, this system does not rely on the traditional dye-quencher-labeled probe, thus reducing the fluorescence background presented in the assay. Furthermore, our one-step detection protocol is performed on-chip at isothermal conditions (37 °C) without using complicated and time-consuming off-chip probe hybridization and denaturation. This miniaturized and fully packed IMPACT chip demonstrates sensitive and accurate DNA detection within 120 min and paves ways to the next-generation point-of-care diagnostics, responding to emerging and deadly pathogen outbreaks.
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spelling pubmed-75941542020-10-30 Integrated Micropillar Polydimethylsiloxane Accurate CRISPR Detection System for Viral DNA Sensing Hass, Kenneth N. Bao, Mengdi He, Qian Liu, Li He, Jiacheng Park, Myeongkee Qin, Peiwu Du, Ke ACS Omega [Image: see text] A fully Integrated Micropillar Polydimethylsiloxane Accurate CRISPR deTection (IMPACT) system is developed for viral DNA detection. This powerful system is patterned with high-aspect-ratio micropillars to enhance reporter probe binding. After surface modification and probe immobilization, the CRISPR-Cas12a/crRNA complex is injected into the fully enclosed microchannel. With the presence of a double-stranded DNA target, the CRISPR enzyme is activated and denatures the single-stranded DNA reporters from the micropillars. This collateral cleavage releases fluorescence reporters into the assay, and the intensity is linearly proportional to the target DNA concentration ranging from 0.1 to 10 nM. Importantly, this system does not rely on the traditional dye-quencher-labeled probe, thus reducing the fluorescence background presented in the assay. Furthermore, our one-step detection protocol is performed on-chip at isothermal conditions (37 °C) without using complicated and time-consuming off-chip probe hybridization and denaturation. This miniaturized and fully packed IMPACT chip demonstrates sensitive and accurate DNA detection within 120 min and paves ways to the next-generation point-of-care diagnostics, responding to emerging and deadly pathogen outbreaks. American Chemical Society 2020-10-13 /pmc/articles/PMC7594154/ /pubmed/33134706 http://dx.doi.org/10.1021/acsomega.0c03917 Text en © 2020 American Chemical Society This is an open access article published under an ACS AuthorChoice License (http://pubs.acs.org/page/policy/authorchoice_termsofuse.html) , which permits copying and redistribution of the article or any adaptations for non-commercial purposes.
spellingShingle Hass, Kenneth N.
Bao, Mengdi
He, Qian
Liu, Li
He, Jiacheng
Park, Myeongkee
Qin, Peiwu
Du, Ke
Integrated Micropillar Polydimethylsiloxane Accurate CRISPR Detection System for Viral DNA Sensing
title Integrated Micropillar Polydimethylsiloxane Accurate CRISPR Detection System for Viral DNA Sensing
title_full Integrated Micropillar Polydimethylsiloxane Accurate CRISPR Detection System for Viral DNA Sensing
title_fullStr Integrated Micropillar Polydimethylsiloxane Accurate CRISPR Detection System for Viral DNA Sensing
title_full_unstemmed Integrated Micropillar Polydimethylsiloxane Accurate CRISPR Detection System for Viral DNA Sensing
title_short Integrated Micropillar Polydimethylsiloxane Accurate CRISPR Detection System for Viral DNA Sensing
title_sort integrated micropillar polydimethylsiloxane accurate crispr detection system for viral dna sensing
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7594154/
https://www.ncbi.nlm.nih.gov/pubmed/33134706
http://dx.doi.org/10.1021/acsomega.0c03917
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