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.
|Publication status||Accepted/In press - 2023|
Bibliographical noteFunding 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.).
© 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)
- Physics and Astronomy(all)