Original Research Paper
A zwitterionic polymer-inspired material mediated efficient CRISPR-Cas9 gene editing

https://doi.org/10.1016/j.ajps.2022.08.001Get rights and content
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Abstract

The type II prokaryotic CRISPR (clustered regularly interspaced short palindromic repeats)/Cas9 (CRISPR/Cas9) adaptive immune system is a cutting-edge genome-editing toolbox. However, its applications are still limited by its inefficient transduction. Herein, we present a novel gene vector, the zwitterionic polymer-inspired material with branched structure (ZEBRA) for efficient CRISPR/Cas9 delivery. Polo-like kinase 1 (PLK1) acts as a master regulator of mitosis and overexpresses in multiple tumor cells. The Cas9 and single guide sgRNA (sgRNA)-encoded plasmid was transduced to knockout Plk1 gene, which was expected to inhibit the expression of PLK1. Our studies demonstrated that ZEBRA enabled to transduce the CRISPR/Cas9 system with large size into the cells efficiently. The transduction with ZEBRA was cell line dependent, which showed ∼10-fold higher in CD44-positive cancer cell lines compared with CD44-negative ones. Furthermore, ZEBRA induced high-level expression of Cas9 proteins by the delivery of CRISPR/Cas9 and efficient gene editing of Plk1 gene, and inhibited the tumor cell growth significantly. This zwitterionic polymer-inspired material is an effective and targeted gene delivery vector and further studies are required to explore its potential in gene delivery applications.

Graphical abstract

We developed a zwitterionic polymer-inspired material with branched structure (ZEBRA) with high bioactivity, high transfection efficiency, specific targeting capability, and high biosafety for CRISPR/Cas9 delivery.

Image, graphical abstract
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Keywords

CRISPR/Cas9
Gene editing
Zwitterionic polymers
CD44
PLK1

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1

These authors contributed to the work equally.