Engineered CRISPR/Cas Ribonucleoproteins for Enhanced Biosensing and Bioimaging

IF 6.7 1区化学 Q1 CHEMISTRY, ANALYTICAL Analytical Chemistry Pub Date : 2025-03-11 DOI:10.1021/acs.analchem.4c06789

Linxin Cao, Zeyuan Wang, Chunyang Lei, Zhou Nie

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Abstract

CRISPR-Cas systems represent a highly programmable and precise nucleic acid-targeting platform, which has been strategically engineered as a versatile toolkit for biosensing and bioimaging applications. Nevertheless, their analytical performance is constrained by inherent functional and activity limitations of natural CRISPR/Cas systems, underscoring the critical role of molecular engineering in enhancing their capabilities. This review comprehensively examines recent advancements in engineering CRISPR/Cas ribonucleoproteins (RNPs) to enhance their functional capabilities for advanced molecular detection and cellular imaging. We explore innovative strategies for developing enhanced CRISPR/Cas RNPs, including Cas protein engineering through protein mutagenesis and fusion techniques, and guide RNA engineering via chemical and structural modifications. Furthermore, we evaluate these engineered RNPs’ applications in sensitive biomarker detection and live-cell genomic DNA and RNA monitoring, while analyzing the current challenges and prospective developments in CRISPR-Cas RNP engineering for advanced biosensing and bioimaging.

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用于增强生物传感和生物成像的工程CRISPR/Cas核糖核蛋白

CRISPR-Cas系统代表了一个高度可编程和精确的核酸靶向平台，它已被战略性地设计为生物传感和生物成像应用的多功能工具包。然而，它们的分析性能受到天然CRISPR/Cas系统固有的功能和活性限制的限制，强调了分子工程在增强其能力方面的关键作用。本文综述了CRISPR/Cas核糖核蛋白（RNPs）工程的最新进展，以增强其在高级分子检测和细胞成像方面的功能能力。我们探索了开发增强型CRISPR/Cas RNPs的创新策略，包括通过蛋白质诱变和融合技术进行Cas蛋白工程，以及通过化学和结构修饰指导RNA工程。此外，我们评估了这些工程RNP在敏感生物标志物检测和活细胞基因组DNA和RNA监测方面的应用，同时分析了CRISPR-Cas RNP工程在先进生物传感和生物成像方面的当前挑战和未来发展。

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来源期刊

Analytical Chemistry 化学-分析化学

CiteScore

12.10

自引率

12.20%

发文量

1949

审稿时长

1.4 months

期刊介绍： Analytical Chemistry, a peer-reviewed research journal, focuses on disseminating new and original knowledge across all branches of analytical chemistry. Fundamental articles may explore general principles of chemical measurement science and need not directly address existing or potential analytical methodology. They can be entirely theoretical or report experimental results. Contributions may cover various phases of analytical operations, including sampling, bioanalysis, electrochemistry, mass spectrometry, microscale and nanoscale systems, environmental analysis, separations, spectroscopy, chemical reactions and selectivity, instrumentation, imaging, surface analysis, and data processing. Papers discussing known analytical methods should present a significant, original application of the method, a notable improvement, or results on an important analyte.