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

Yagiz Alp Aksoy; Biyao Yang; Wenjie Chen; Tzongtyng Hung; Rhiannon P Kuchel; Nathan W Zammit; Shane T Grey; Ewa M Goldys; Wei Deng

doi:10.1021/acsami.0c16380

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

ACS Appl Mater Interfaces. 2020 Nov 25;12(47):52433-52444. doi: 10.1021/acsami.0c16380. Epub 2020 Nov 11.

Authors

Yagiz Alp Aksoy^{1

2

3}, Biyao Yang⁴, Wenjie Chen⁵, Tzongtyng Hung⁶, Rhiannon P Kuchel⁷, Nathan W Zammit^{8

9}, Shane T Grey^{8

9}, Ewa M Goldys⁴, Wei Deng⁴

Affiliations

¹ Diabetes and Metabolism Division, Garvan Institute of Medical Research, Sydney, NSW 2010, Australia.
² Department of Biomedical Sciences, Faculty of Medicine and Health Sciences, Centre for Motor Neuron Disease Research, Macquarie University, Sydney, NSW 2109, Australia.
³ Sydney Medical School, The University of Sydney, Sydney, NSW 2006, Australia.
⁴ ARC Centre of Excellence for Nanoscale Biophotonics, Graduate School of Biomedical Engineering, University of New South Wales, Sydney, NSW 2052, Australia.
⁵ Center for Pharmaceutical Engineering and Sciences, Department of Pharmaceutics, School of Pharmacy, Virginia Commonwealth University, Richmond, Virginia 23298, United States.
⁶ The Biological Resource Imaging Laboratory, University of New South Wales, Sydney, NSW 2052, Australia.
⁷ Electron Microscope Unit, University of New South Wales, Sydney, NSW 2052, Australia.
⁸ Garvan Institute of Medical Research, Sydney, NSW 2010, Australia.
⁹ Faculty of Medicine, University of New South Wales, Sydney, NSW 2052, Australia.

PMID: 33174413
DOI: 10.1021/acsami.0c16380

Abstract

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.

Keywords: CRISPR-Cas9; light triggering; liposomes; protein release; temporal control.

MeSH terms

Animals
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

Substances

Liposomes
Tumor Necrosis Factor-alpha
enhanced green fluorescent protein
Green Fluorescent Proteins
Singlet Oxygen
TNFAIP3 protein, human
Tumor Necrosis Factor alpha-Induced Protein 3