Stimuli-responsive peptide nanocarriers for tumor-specific CRISPR/Cas9 delivery and precision genome editing

IF 9.4 1区化学 Q1 CHEMISTRY, PHYSICAL

Journal of Colloid and Interface Science Pub Date : 2025-05-19 DOI:10.1016/j.jcis.2025.137932

Rui Wang , Yujie Yang , Zizai Wang , Chen Ma , Mengqi Wu , Yangyang Du , Xiaoming Zhang , Meiwen Cao , Hai Xu

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引用次数: 0

Abstract

CRISPR/Cas9 ribonucleoprotein (RNP) delivery remains a critical challenge due to its large size, instability, and off-target effects. Here, we report a stimuli-responsive cationic amphiphilic peptide, (CR₃)₃C, designed for cancer-targeted delivery of CRISPR/Cas9 RNP. The peptide integrates three functional domains: (1) a naphthyl-diphenylalanine (Nap-FF) motif enabling self-assembly into stable nanoparticles via aromatic interactions, (2) a matrix metalloproteinase-7 (MMP7)-cleavable linker (GPLGLA) for tumor microenvironment-specific activation, and (3) a redox-responsive cationic domain ((CRRR)₃-C) for electrostatic RNP binding and glutathione (GSH)-triggered intracellular release. The (CR₃)₃C/RNP nanocomplexes (108.8 nm diameter, ζ = +10.89 mV) demonstrate exceptional stability and cellular uptake efficiency. Mechanistic studies reveal caveolae-mediated endocytosis and lipid raft-associated pathways, proton sponge effect-driven endosomal escape, and nuclear localization facilitated by Cas9′s nuclear localization signal. In HeLa-EGFP cells, (CR₃)₃C/RNP shows 33.8 % gene editing efficiency at 100 nM RNP with >90 % cell viability. This work establishes a programmable, non-viral platform that synergizes enzymatic and redox responsiveness for tumor-targeted genome editing, addressing critical barriers in CRISPR therapeutics.

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用于肿瘤特异性CRISPR/Cas9传递和精确基因组编辑的刺激响应肽纳米载体

由于CRISPR/Cas9核糖核蛋白（RNP）的大尺寸、不稳定性和脱靶效应，其递送仍然是一个关键的挑战。在这里，我们报道了一种刺激反应性阳离子两亲性肽（CR3）3C，设计用于靶向递送CRISPR/Cas9 RNP。该肽整合了三个功能域：(1)一个萘基二苯丙氨酸（Nap-FF）基序，能够通过芳香相互作用自组装成稳定的纳米颗粒；(2)一个基质金属蛋白酶-7 (MMP7)-可切割连接体（GPLGLA），用于肿瘤微环境特异性激活；(3)一个氧化还原响应阳离子结构域（CRRR）3- c，用于静电RNP结合和谷胱甘肽（GSH）触发的细胞内释放。（CR3）3C/RNP纳米复合物（直径108.8 nm, ζ = +10.89 mV）表现出优异的稳定性和细胞摄取效率。机制研究揭示了小泡介导的内吞作用和脂筏相关途径，质子海绵效应驱动的内体逃逸，以及Cas9的核定位信号促进的核定位。在HeLa-EGFP细胞中，（CR3）3C/RNP在100 nM RNP下的基因编辑效率为33.8%，细胞存活率为90%。这项工作建立了一个可编程的非病毒平台，协同酶和氧化还原反应，用于肿瘤靶向基因组编辑，解决CRISPR治疗中的关键障碍。

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来源期刊

Journal of Colloid and Interface Science 化学-物理化学

CiteScore

16.10

自引率

7.10%

发文量

2568

审稿时长

2 months

期刊介绍： The Journal of Colloid and Interface Science publishes original research findings on the fundamental principles of colloid and interface science, as well as innovative applications in various fields. The criteria for publication include impact, quality, novelty, and originality. Emphasis: The journal emphasizes fundamental scientific innovation within the following categories: A.Colloidal Materials and Nanomaterials B.Soft Colloidal and Self-Assembly Systems C.Adsorption, Catalysis, and Electrochemistry D.Interfacial Processes, Capillarity, and Wetting E.Biomaterials and Nanomedicine F.Energy Conversion and Storage, and Environmental Technologies