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Scaffold-mediated non-viral delivery platform for CRISPR/Cas9-based genome editing
Jiah Shin Chin1, Wai Hon Chooi2, Hongxia Wang3
1School of Chemical and Biomedical Engineering, Nanyang Technological University, Singapore 637459, Singapore; NTU Institute of Health Technologies, Interdisciplinary Graduate School, Nanyang Technological University, Singapore 639798, Singapore.
Acta Biomaterialia
|April 13, 2019
Summary
This study presents a novel scaffold for localized, non-viral CRISPR-Cas9 delivery, enhancing tissue regeneration. The mussel-inspired coating ensures efficient gene editing with reduced off-target effects.
Area of Science:
- Biotechnology
- Regenerative Medicine
- Molecular Biology
Background:
- CRISPR-Cas9 genome editing is crucial for controlling cell fate in tissue regeneration.
- Existing non-viral delivery methods often use micro-/nano-particles, risking systemic off-target effects.
- Localized and sustained delivery is needed for in vivo applications.
Purpose of the Study:
- To develop a novel scaffold for localized, non-viral delivery of CRISPR-Cas9 components.
- To improve the efficiency and safety of genome editing for tissue regeneration.
Main Methods:
- Developed bio-mimicking fiber scaffolds coated with polyDOPA-melanin (pDOPA) for bioadhesion.
- Adsorbed Cas9:sgRNA lipofectamine complexes onto the pDOPA-coated scaffolds.
- Utilized U2OS.EGFP cells to evaluate genome-editing efficiency and cell viability.
Main Results:
- pDOPA coating enhanced loading efficiency of Cas9:sgRNA complexes.
- Laminin coating maintained cell viability and proliferation for effective gene editing.
- Achieved 21.5% gene editing efficiency via reverse transfection from the scaffold.
Conclusions:
- Demonstrated the efficacy of fiber scaffolds for localized, sustained, non-viral CRISPR-Cas9 delivery.
- This platform shows potential for applications in tissue regeneration.
- The localized delivery minimizes risks associated with systemic administration.
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