CRISPR-Enhanced Hydrogel Microparticles for Multiplexed Detection of Nucleic Acids
© 2023 The Authors. Advanced Science published by Wiley-VCH GmbH..
CRISPR/Cas systems offer a powerful sensing mechanism to transduce sequence-specific information into amplified analytical signals. However, performing multiplexed CRISPR/Cas assays remains challenging and often requires complex approaches for multiplexed assays. Here, a hydrogel-based CRISPR/Cas12 system termed CLAMP (Cas-Loaded Annotated Micro-Particles) is described. The approach compartmentalizes the CRISPR/Cas reaction in spatially-encoded hydrogel microparticles (HMPs). Each HMP is identifiable by its face code and becomes fluorescent when target DNA is present. The assay is further streamlined by capturing HMPs inside a microfluidic device; the captured particles are then automatically recognized by a machine-learning algorithm. The CLAMP assay is fast, highly sensitive (attomolar detection limits with preamplification), and capable of multiplexing in a single-pot assay. As a proof-of-concept clinical application, CLAMP is applied to detect nucleic acid targets of human papillomavirus in cervical brushing samples.
| Medienart: |
E-Artikel |
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| Erscheinungsjahr: |
2023 |
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| Erschienen: |
2023 |
| Enthalten in: |
Zur Gesamtaufnahme - volume:10 |
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| Enthalten in: |
Advanced science (Weinheim, Baden-Wurttemberg, Germany) - 10(2023), 10 vom: 20. Apr., Seite e2206872 |
| Sprache: |
Englisch |
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| Beteiligte Personen: |
Roh, Yoon Ho [VerfasserIn] |
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| Links: |
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| Anmerkungen: |
Date Completed 06.04.2023 Date Revised 11.04.2023 published: Print-Electronic Citation Status MEDLINE |
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| doi: |
10.1002/advs.202206872 |
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| funding: |
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| Förderinstitution / Projekttitel: |
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| PPN (Katalog-ID): |
NLM352356189 |
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| 520 | |a © 2023 The Authors. Advanced Science published by Wiley-VCH GmbH. | ||
| 520 | |a CRISPR/Cas systems offer a powerful sensing mechanism to transduce sequence-specific information into amplified analytical signals. However, performing multiplexed CRISPR/Cas assays remains challenging and often requires complex approaches for multiplexed assays. Here, a hydrogel-based CRISPR/Cas12 system termed CLAMP (Cas-Loaded Annotated Micro-Particles) is described. The approach compartmentalizes the CRISPR/Cas reaction in spatially-encoded hydrogel microparticles (HMPs). Each HMP is identifiable by its face code and becomes fluorescent when target DNA is present. The assay is further streamlined by capturing HMPs inside a microfluidic device; the captured particles are then automatically recognized by a machine-learning algorithm. The CLAMP assay is fast, highly sensitive (attomolar detection limits with preamplification), and capable of multiplexing in a single-pot assay. As a proof-of-concept clinical application, CLAMP is applied to detect nucleic acid targets of human papillomavirus in cervical brushing samples | ||
| 650 | 4 | |a Journal Article | |
| 650 | 4 | |a Research Support, N.I.H., Extramural | |
| 650 | 4 | |a Research Support, Non-U.S. Gov't | |
| 650 | 4 | |a CRISPR/Cas | |
| 650 | 4 | |a human papillomavirus | |
| 650 | 4 | |a hydrogel microparticles | |
| 650 | 4 | |a isothermal amplification | |
| 650 | 4 | |a multiplexed assays | |
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| 700 | 1 | |a Lee, Chang Yeol |e verfasserin |4 aut | |
| 700 | 1 | |a Lee, Sujin |e verfasserin |4 aut | |
| 700 | 1 | |a Kim, Hyunho |e verfasserin |4 aut | |
| 700 | 1 | |a Ly, Amy |e verfasserin |4 aut | |
| 700 | 1 | |a Castro, Cesar M |e verfasserin |4 aut | |
| 700 | 1 | |a Cheon, Jinwoo |e verfasserin |4 aut | |
| 700 | 1 | |a Lee, Jae-Hyun |e verfasserin |4 aut | |
| 700 | 1 | |a Lee, Hakho |e verfasserin |4 aut | |
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