Department of Chemistry, University of Massachusetts , 710 North Pleasant Street, Amherst, Massachusetts 01003, United States.
ACS Nano. 2017 Mar 28;11(3):2452-2458. doi: 10.1021/acsnano.6b07600. Epub 2017 Jan 31.
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.
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。