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https://hdl.handle.net/10356/142907
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DC Field | Value | Language |
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dc.contributor.author | Chin, Jiah Shin | en_US |
dc.contributor.author | Chooi, Wai Hon | en_US |
dc.contributor.author | Wang, Hongxia | en_US |
dc.contributor.author | Ong, William | en_US |
dc.contributor.author | Leong, Kam W. | en_US |
dc.contributor.author | Chew, Sing Yian | en_US |
dc.date.accessioned | 2020-07-08T03:19:08Z | - |
dc.date.available | 2020-07-08T03:19:08Z | - |
dc.date.issued | 2019 | - |
dc.identifier.citation | 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 | en_US |
dc.identifier.issn | 1742-7061 | en_US |
dc.identifier.uri | https://hdl.handle.net/10356/142907 | - |
dc.description.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. | en_US |
dc.language.iso | en | en_US |
dc.relation.ispartof | Acta Biomaterialia | en_US |
dc.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. | en_US |
dc.subject | Engineering::Bioengineering | en_US |
dc.title | Scaffold-mediated non-viral delivery platform for CRISPR/Cas9-based genome editing | en_US |
dc.type | Journal Article | en |
dc.contributor.school | School of Chemical and Biomedical Engineering | en_US |
dc.contributor.school | Interdisciplinary Graduate School (IGS) | en_US |
dc.contributor.school | Lee Kong Chian School of Medicine (LKCMedicine) | en_US |
dc.contributor.organization | NTU Institute of Health Technologies | en_US |
dc.identifier.doi | 10.1016/j.actbio.2019.04.020 | - |
dc.description.version | Accepted version | en_US |
dc.identifier.volume | 90 | en_US |
dc.identifier.spage | 60 | en_US |
dc.identifier.epage | 70 | en_US |
dc.subject.keywords | Gene Delivery | en_US |
dc.subject.keywords | Cas9 Protein | en_US |
item.fulltext | With Fulltext | - |
item.grantfulltext | open | - |
Appears in Collections: | SCBE Journal Articles |
Files in This Item:
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Figure 1.tif | 45.91 MB | TIFF | View/Open | |
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 | |
Figure captions_revised.docx | 13.38 kB | Microsoft Word | View/Open | |
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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