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


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
JournalAdvanced Science
Publication statusAccepted/In press - 2023

Bibliographical note

Funding Information:
Y.H.R. and C.Y.L. contributed equally to this work. NIH R21DA049577 (H.L.), R01CA229777 (H.L.), R01CA239078 (H.L.), R01CA237500 (H.L.), R21CA267222 (H.L.), U01CA233360 (H.L., C.M.C.), R01CA264363 (C.M.C, H.L.), MGH Scholar Fund (H.L); Robert Wood Johnson Foundation (C.M.C.); National Research Foundation (Republic of Korea) NRF‐2021R1A6A3A14044562 (H.K.); Institute for Basic Science (Korea) IBS‐R026‐D1 (J.‐H.L, J.C.).

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

All Science Journal Classification (ASJC) codes

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


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