Hybridization Chain Reaction (HCR) is a DNA-based target-induced cascade reaction. Due to its unique enzyme-free amplification feature, HCR is often employed for sensing applications. Despite its standalone capabilities, HCR is highly modular, therefore it can be repurposed when coupled with latest technological advancements. To this effect, we have developed a detection approach which combines the signal amplification feature of HCR with the programmability and sensitivity of CRISPR-Cas12a system. By incorporating CRISPR-Cas12a, we have achieved remarkable sensitivity, and the ability to rapidly reprogram the assay against a different target by merely changing a single reaction component. We have demonstrated this programmability by employing the assay for two distinct targets, Hepatitis B virus, and Tobacco Curly Shoot virus. Furthermore, this assay has enabled the detection of both single and double stranded DNA targets using gel electrophoresis and fluorescence for signal readout.

Hybridization chain reaction and CRISPR-Cas systems for disease diagnostics

ACS Spring 2022

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