CRISPR-Enhanced Hydrogel Microparticles for Multiplexed Detection of Nucleic Acids

Yoon Ho Roh, Chang Yeol Lee, Sujin Lee, Hyunho Kim, Amy Ly, Cesar M. Castro, Jinwoo Cheon, Jae Hyun Lee, Hakho Lee

Research output: Contribution to journalArticlepeer-review

12 Citations (Scopus)

Abstract

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.

Original languageEnglish
Article number2206872
JournalAdvanced Science
Volume10
Issue number10
DOIs
Publication statusPublished - 2023 Apr 5

Bibliographical note

Publisher Copyright:
© 2023 The Authors. Advanced Science published by Wiley-VCH GmbH.

All Science Journal Classification (ASJC) codes

  • Medicine (miscellaneous)
  • General Chemical Engineering
  • General Materials Science
  • Biochemistry, Genetics and Molecular Biology (miscellaneous)
  • General Engineering
  • General Physics and Astronomy

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