Enhanced CRISPR‐cas systems for genome editing and molecular diagnostics
TL;DR
The current developments in engineering of Cas effector protein, optimizing guide RNA designs, developing advanced reporter probes, and utilizing chemical additives are summarized and their significant contribution to increasing the fidelity of editing and diagnostic sensitivity is highlighted.
Abstract
CRISPR‐Cas systems have far‐reaching implications for genome editing and molecular diagnostics, and are emerging as important tools in biomedical research and clinical practice. These systems enable precise genetic manipulation and highly sensitive nucleic acid detection, capabilities that are invaluable for therapeutic development and pathogen surveillance. Nevertheless, native CRISPR‐Cas systems possess inherent limitations that constrain their broader utility. These limitations in genome editing include insufficient editing efficiency, a restricted range of targets due to protospacer adjacent motif constraints and non‐negligible off‐target effects. In molecular diagnostics, the remaining problems include detecting low‐abundance targets, achieving high specificity, and ensuring effective compatibility with isothermal amplification. To address these limitations, researchers have proposed many enhancement strategies that mainly center around four aspects: engineering of Cas effector protein, optimizing guide RNA designs, developing advanced reporter probes, and utilizing chemical additives. This review summarizes the current developments in these areas and highlights their significant contribution to increasing the fidelity of editing and diagnostic sensitivity. In addition, we discuss future prospects for enhanced CRISPR‐Cas systems, focusing on their accelerating clinical translation and expanding biomedical applications.