# Advances in nanozyme-assisted CRISPR diagnostic technology

## TL;DR

This review explores how nanozymes can enhance CRISPR diagnostics by improving signal output and reducing reliance on external enzymes.

## Contribution

The paper provides a comprehensive evaluation of nanozyme-assisted CRISPR diagnostics and their integration mechanisms.

## Key findings

- Nanozymes improve the catalytic efficiency and stability of CRISPR diagnostics.
- Integration of nanozymes enables signal amplification and detection of both nucleic and non-nucleic acid targets.
- Challenges include complex separation and slow detection rates, with future directions focusing on homogenous systems and integrated devices.

## Abstract

The clustered regularly interspaced short palindromic repeats (CRISPR) and CRISPR-associated proteins (Cas) system has significant potential in biological diagnostics because of its precise nucleic acid identification abilities. Traditional CRISPR diagnostics, however, have limitations such as insufficient signal output, dependence on exogenous enzymes, and high equipment demands. Nanozymes, as nanomaterials with enzyme-mimetic catalytic activity, integrate the catalytic efficiency of natural enzymes with the stability and modifiability of nanomaterials, providing a viable resolution to the limitations in CRISPR diagnostics. This article comprehensively evaluates the advancements in nanozyme-enhanced CRISPR diagnostic technologies. Furthermore, it delineates the fundamental attributes of the CRISPR diagnostic system and nanozymes, as well as the necessity of their integration. Moreover, the coupling mechanisms between the CRISPR/Cas system and nanozymes, including the regulation of nanozyme catalytic activity by Cas protein function and CRISPR signal amplification facilitated by nanozymes, were also comprehensively evaluated. The application of this technique in detecting nucleic acid and non-nucleic acid targets was assessed. Further, this study discusses the current limitations of this technology, such as complex separation of heterogeneous systems, laborious reaction protocols, and slow detection rates. The future advancements, such as the establishment of homogenous systems, the creation of integrated devices, and the utilization of single-atom nanozymes, have also been discussed in this review. The results of this study will provide references for the comprehensive integration of nanozymes and CRISPR technology, together with their diagnostic applications.