A dendrimer enhanced CRISPR/Cas12a performance for the evaluation of acidogenic capacity in cariogenic bacteria

Abstract

Dental caries, a highly prevalent global health issue, is fundamentally driven by the acidogenic metabolism of cariogenic bacteria such as Streptococcus mutans. Conventional approaches for assessing acidogenic capacity remain limited by complex operational requirements and insufficient sensitivity for point-of-care use. Herein, we engineer a CRISPR/Cas12a-powered dendritic DNA nanostructure via hybridization chain reaction to construct a highly efficient signal amplification system. The designed activator strands self-assemble into dendritic nano-assemblies that enable multivalent immobilization of Cas12a-crRNA complexes through programmable base pairing, resulting in a confined microdomain with dramatically enhanced local enzyme density and accelerated trans-cleavage kinetics. The CRISPR/Cas dendritic aggregate enables high-density immobilization of Cas12a, resulting in increased local enzyme concentration and enhanced catalytic cleavage efficiency. By integrating this system with a smart pH-responsive molecular release mechanism, we further develop a paper-based origami biosensor (CDEOB) for the on-site and equipment-free quantification of acidogenic pathogens. The biosensor achieves an ultra-low detection limit of 98.1 CFU/mL in saliva and demonstrates excellent reliability across 101 clinical samples without pre-amplification. This work provides a novel functional nucleic acid material design strategy and a versatile biosensing platform, highlighting substantial potential for early diagnosis of dental caries and other infectious diseases at the point of care.

Keywords

acidogenic capacity / cariogenic bacteria / CRISPR/Cas12a / dendrimer / performance

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