Spatial and Temporal Control of CRISPR-Cas9-Mediated Gene Editing Delivered via a Light-Triggered Liposome System

• Published in: ACS applied materials & interfaces 2020-11, Vol.12 (47), p.52433-52444
• Main Authors: Aksoy, Yagiz Alp, Yang, Biyao, Chen, Wenjie, Hung, Tzongtyng, Kuchel, Rhiannon P, Zammit, Nathan W, Grey, Shane T, Goldys, Ewa M, Deng, Wei
• Format: Article
• Language: English
• Subjects: Animals; Biological and Medical Applications of Materials and Interfaces; CRISPR-Cas Systems - genetics; Embryo, Nonmammalian - metabolism; Gene Editing - methods; Gene Expression - drug effects; Genes, Reporter; Green Fluorescent Proteins - genetics; Green Fluorescent Proteins - metabolism; HEK293 Cells; Humans; Light; Liposomes - chemistry; Singlet Oxygen - metabolism; Tumor Necrosis Factor alpha-Induced Protein 3 - genetics; Tumor Necrosis Factor alpha-Induced Protein 3 - metabolism; Tumor Necrosis Factor-alpha - pharmacology; Zebrafish - growth & development; Zebrafish - metabolism
• ISSN: 1944-8244; 1944-8252
• DOI: 10.1021/acsami.0c16380

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 in vivo 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.