Please use this identifier to cite or link to this item:
https://hdl.handle.net/10356/142907
Title: | Scaffold-mediated non-viral delivery platform for CRISPR/Cas9-based genome editing | Authors: | Chin, Jiah Shin Chooi, Wai Hon Wang, Hongxia Ong, William Leong, Kam W. Chew, Sing Yian |
Keywords: | Engineering::Bioengineering | Issue Date: | 2019 | Source: | Chin, J. S., Chooi, W. H., Wang, H., Ong, W., Leong, K. W., & Chew, S. Y. (2019). Scaffold-mediated non-viral delivery platform for CRISPR/Cas9-based genome editing. Acta Biomaterialia, 90, 60-70. doi:10.1016/j.actbio.2019.04.020 | Journal: | Acta Biomaterialia | Abstract: | Genome editing, especially via the simple and versatile type II CRISPR/Cas9 system, offers an effective avenue to precisely control cell fate, an important aspect of tissue regeneration. Unfortunately, most CRISPR/Cas9 non-viral delivery strategies only utilise micro-/nano-particle delivery methods. While these approaches provide reasonable genomic editing efficiencies, their systemic delivery may lead to undesirable off-target effects. For in vivo applications, a more localized and sustained delivery approach may be useful, particularly in the context of tissue regeneration. Here, we developed a scaffold that delivers the CRISPR/Cas9 components (i.e. single guide RNA (sgRNA) and Cas9 protein complexes) in a localized and non-viral manner. Specifically, using mussel-inspired bioadhesive coating, polyDOPA-melanin (pDOPA), we absorbed Cas9:sgRNA lipofectamine complexes onto bio-mimicking fiber scaffolds. To evaluate the genome-editing efficiency of this platform, U2OS.EGFP cells were used as the model cell type. pDOPA coating was essential in allowing Cas9:sgRNA lipofectamine complexes to adhere onto the scaffolds with a higher loading efficiency, while laminin coating was necessary for maintaining cell viability and proliferation on the pDOPA-coated fibers for effective gene editing (21.5% editing efficiency, p < 0.001). Importantly, U2OS.EGFP cells took up Cas9:sgRNA lipofectamine complexes directly from the scaffolds via reverse transfection. Overall, we demonstrate the efficacy of such fiber scaffolds in providing localized, sustained and non-viral delivery of Cas9:sgRNA complexes. Such genome editing scaffolds may find useful applications in tissue regeneration. | URI: | https://hdl.handle.net/10356/142907 | ISSN: | 1742-7061 | DOI: | 10.1016/j.actbio.2019.04.020 | Rights: | © 2019 Acta Materialia Inc. All rights reserved. This paper was published by Elsevier Ltd. in Acta Biomaterialia and is made available with permission of Acta Materialia Inc. | Fulltext Permission: | open | Fulltext Availability: | With Fulltext |
Appears in Collections: | SCBE Journal Articles |
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Figure 2.tif | 31.65 MB | TIFF | View/Open | |
Figure 3.tif | 64.65 MB | TIFF | View/Open | |
Figure 4.tif | 26.18 MB | TIFF | View/Open | |
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Scaffold-mediated non-viral ...pdf | 396.2 kB | Adobe PDF | View/Open | |
Supplementary-Scaffold-mediated non-viral ...pdf | 849.97 kB | Adobe PDF | View/Open |
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