Diabetes and Metabolism Division, Garvan Institute of Medical Research, Sydney, NSW 2010, Australia.
Department of Biomedical Sciences, Faculty of Medicine and Health Sciences, Centre for Motor Neuron Disease Research, Macquarie University, Sydney, NSW 2109, Australia.
ACS Appl Mater Interfaces. 2020 Nov 25;12(47):52433-52444. doi: 10.1021/acsami.0c16380. Epub 2020 Nov 11.
The CRISPR-Cas9 and related systems offer a unique genome-editing tool allowing facile and efficient introduction of heritable and locus-specific sequence modifications in the genome. Despite its molecular precision, temporal and spatial control of gene editing with the CRISPR-Cas9 system is very limited. We developed a light-sensitive liposome delivery system that offers a high degree of spatial and temporal control of gene editing with the CRISPR-Cas9 system. We demonstrated its efficient protein release by respectively assessing the targeted knockout of the eGFP gene in human HEK293/GFP cells and the TNFAIP3 gene in TNFα-induced HEK293 cells. We further validated our results at a single-cell resolution using an eGFP reporter system in zebrafish (77% knockout). These findings indicate that light-triggered liposomes may have new options for precise control of CRISPR-Cas9 release and editing.
CRISPR-Cas9 系统和相关系统提供了一种独特的基因组编辑工具，可轻松有效地在基因组中引入可遗传和特定位置的序列修饰。尽管具有分子精度，但 CRISPR-Cas9 系统的基因编辑的时空控制非常有限。我们开发了一种光敏感脂质体递药系统，可实现 CRISPR-Cas9 系统的高度时空基因编辑控制。我们通过分别评估人 HEK293/GFP 细胞中 eGFP 基因的靶向敲除和 TNFα 诱导的 HEK293 细胞中 TNFAIP3 基因的靶向敲除，证明了其高效的蛋白释放。我们进一步使用斑马鱼中的 eGFP 报告系统（77%的敲除率）在单细胞分辨率上验证了我们的结果。这些发现表明，光触发脂质体可能为 CRISPR-Cas9 释放和编辑的精确控制提供了新的选择。
