Direct cellular delivery of CRISPR/Cas9 complexes is of great significance for genome editing and other recently developed applications, such as gene expression regulation and RNA/DNA imaging. Here, we first constructed a graphene oxide (GO)-polyethylene glycol (PEG)-polyethylenimine (PEI) nanocarrier for the delivery of high-molecular-weight Cas9/single-guide RNA (sgRNA) complexes for endocytosis, endosomal escape, nuclear entry, and gene editing. The results demonstrate that the nanocarrier can be used successfully for efficient gene editing in human AGS cells with an efficiency of ∼39%. The results also show that this nanocarrier can protect sgRNA from enzymatic degradation, thus exhibiting extremely high stability, which is critical for future in vivo applications. Thus, this GO-mediated Cas9/sgRNA delivery system has potential as a new approach for biomedical research and targeted gene engineering applications.

中文翻译：

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氧化石墨烯介导的Cas9 / sgRNA传递可实现有效的基因组编辑†

CRISPR / Cas9复合物的直接细胞递送对基因组编辑和其他最近开发的应用（如基因表达调控和RNA / DNA成像）具有重要意义。在这里，我们首先构建了氧化石墨烯（GO）-聚乙二醇（PEG）-聚乙烯亚胺（PEI）纳米载体，用于递送高分子量Cas9 /单向导RNA（sgRNA）复合物，用于内吞，内体逃逸，核进入，以及基因编辑。结果表明，纳米载体可以成功地用于人AGS细胞的有效基因编辑，效率约为39％。结果还表明，这种纳米载体可以保护sgRNA免受酶促降解，因此显示出极高的稳定性，这对于将来在体内的应用至关重要应用程序。因此，这种GO介导的Cas9 / sgRNA递送系统具有作为生物医学研究和靶向基因工程应用的新方法的潜力。