直接细胞质递送 CRISPR/Cas9 核糖核蛋白进行高效基因编辑。
Direct Cytosolic Delivery of CRISPR/Cas9-Ribonucleoprotein for Efficient Gene Editing.
机构信息
Department of Chemistry, University of Massachusetts , 710 North Pleasant Street, Amherst, Massachusetts 01003, United States.
出版信息
ACS Nano. 2017 Mar 28;11(3):2452-2458. doi: 10.1021/acsnano.6b07600. Epub 2017 Jan 31.
Abstract
Genome editing through the delivery of CRISPR/Cas9-ribonucleoprotein (Cas9-RNP) reduces unwanted gene targeting and avoids integrational mutagenesis that can occur through gene delivery strategies. Direct and efficient delivery of Cas9-RNP into the cytosol followed by translocation to the nucleus remains a challenge. Here, we report a remarkably highly efficient (∼90%) direct cytoplasmic/nuclear delivery of Cas9 protein complexed with a guide RNA (sgRNA) through the coengineering of Cas9 protein and carrier nanoparticles. This construct provides effective (∼30%) gene editing efficiency and opens up opportunities in studying genome dynamics.
摘要
通过递送 CRISPR/Cas9-核糖核蛋白（Cas9-RNP）进行基因组编辑可减少不必要的基因靶向，并避免通过基因传递策略发生的整合诱变。Cas9-RNP 直接有效地递送到细胞质中，然后转运到细胞核仍然是一个挑战。在这里，我们通过对 Cas9 蛋白和载体纳米颗粒的共同工程化，报告了一种通过共工程化 Cas9 蛋白和载体纳米颗粒，将 Cas9 蛋白复合物与向导 RNA（sgRNA）进行高效（约 90%）直接细胞质/核内递送至细胞质/核内的方法。该构建体提供了有效的（约 30%）基因编辑效率，并为研究基因组动力学开辟了机会。
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