Endosomal Escape and Delivery of CRISPR/Cas9 Genome Editing Machinery Enabled by Nanoscale Zeolitic Imidazolate Framework
Endosomal Escape and Delivery of CRISPR/Cas9 Genome Editing Machinery Enabled by Nanoscale Zeolitic Imidazolate Framework
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DOI：
10.1021/jacs.7b11754
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发表时间：
2018-01-10
影响因子：
15
通讯作者：
Khashab, Niveen M.
中科院分区：
文献类型：
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作者：
Alsaiari, Shahad K.;Patil, Sachin;Khashab, Niveen M.
CRISPR/Cas9是一种组合蛋白（Cas9）和工程单导RNA （sgRNA）基因组编辑平台，为遗传性疾病提供了革命性的解决方案。然而，由于蛋白质的大尺寸和高电荷的RNA成分，它有双重递送问题。在这项工作中，我们报道了第一个由纳米级沸沸体咪唑框架（ZIFs）封装的CRISPR/Cas9的例子，其负载效率为17%，并且质子化的咪唑部分促进了内体逃逸。ZIF-8封装的CRISPR/Cas9 （CC-ZIFs）的基因编辑潜力通过在4天内将绿色荧光蛋白的基因表达降低37%进一步得到验证。纳米级cc - zif具有生物相容性，易于扩展，具有出色的负载能力和完整Cas9蛋白和sgRNA的可控共递送。
CRISPR/Cas9 is a combined protein (Cas9) and an engineered single guide RNA (sgRNA) genome editing platform that offers revolutionary solutions to genetic diseases. It has, however, a double delivery problem owning to the large protein size and the highly charged RNA component. In this work, we report the first example of CRISPR/Cas9 encapsulated by nanoscale zeolitic imidazole frameworks (ZIFs) with a loading efficiency of 17% and enhanced endosomal escape promoted by the protonated imidazole moieties. The gene editing potential of CRISPR/Cas9 encapsulated by ZIF-8 (CC-ZIFs) is further verified by knocking down the gene expression of green fluorescent protein by 37% over 4 days. The nanoscale CC-ZIFs are biocompatible and easily scaled-up offering excellent loading capacity and controlled codelivery of intact Cas9 protein and sgRNA.