CRISPR‐Enhanced Hydrogel Microparticles for Multiplexed Detection of Nucleic Acids

Abstract

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.

Keywords

• CRISPR/Cas
• human papillomavirus
• hydrogel microparticles
• isothermal amplification
• multiplexed assays