氮掺杂碳点促进 CRISPR/Cas 减少环境中的抗生素耐药基因。

Nitrogen-Doped Carbon Dots Facilitate CRISPR/Cas for Reducing Antibiotic Resistance Genes in the Environment.

机构信息

Institute of Environmental Processes and Pollution Control, and School of Environment and Ecology, Jiangnan University, Wuxi 214122, China.

Jiangsu Engineering Laboratory for Biomass Energy and Carbon Reduction Technology, Wuxi 214122, China.

出版信息

J Agric Food Chem. 2024 Feb 21;72(7):3397-3405. doi: 10.1021/acs.jafc.3c08558. Epub 2024 Feb 9.

DOI:10.1021/acs.jafc.3c08558

PMID:38335532

Abstract

The continued acquisition and propagation of antibiotic resistance genes (ARGs) in the environment confound efforts to manage the global rise in antibiotic resistance. Here, CRISPR-Cas9/sgRNAs carried by nitrogen-doped carbon dots (NCDs) were developed to precisely target multi-"high-risk" ARGs (, , and ) commonly detected in the environment. NCDs facilitated the delivery of Cas9/sgRNAs to () without cytotoxicity, achieving sustained elimination of target ARGs. The elimination was optimized using different weight ratios of NCDs and Cas9 protein (1:1, 1:20, and 1:40), and Cas9/multi sgRNAs were designed to achieve multi-cleavage of ARGs in either a single strain or mixed populations. Importantly, NCDs successfully facilitated Cas9/multi sgRNAs for resensitization of antibiotic-resistant bacteria in soil (approaching 50%), whereas Cas9/multi sgRNAs alone were inactivated in the complex environment. This work highlights the potential of a fast and precise strategy to minimize the reservoir of antibiotic resistance in agricultural system.

摘要

在环境中持续获取和传播抗生素耐药基因（ARGs），这给全球抗生素耐药性的上升带来了挑战。在这里，我们开发了由氮掺杂碳点（NCDs）携带的 CRISPR-Cas9/sgRNAs，以精确靶向环境中常见的多“高风险”ARGs（,, 和）。NCDs 促进了 Cas9/sgRNAs 递送到（）而没有细胞毒性，从而实现了靶标 ARGs 的持续消除。通过使用不同重量比的 NCDs 和 Cas9 蛋白（1:1、1:20 和 1:40）优化了消除效果，并设计了 Cas9/多 sgRNAs 以实现 ARGs 在单一菌株或混合种群中的多切割。重要的是，NCDs 成功地促进了 Cas9/多 sgRNAs 用于恢复土壤中抗生素耐药细菌的敏感性（接近 50%），而 Cas9/多 sgRNAs 单独在复杂环境中失活。这项工作强调了一种快速而精确的策略的潜力，该策略可以最大限度地减少农业系统中抗生素耐药性的储存。