Please use this identifier to cite or link to this item: https://hdl.handle.net/10356/142907
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dc.contributor.authorChin, Jiah Shinen_US
dc.contributor.authorChooi, Wai Honen_US
dc.contributor.authorWang, Hongxiaen_US
dc.contributor.authorOng, Williamen_US
dc.contributor.authorLeong, Kam W.en_US
dc.contributor.authorChew, Sing Yianen_US
dc.date.accessioned2020-07-08T03:19:08Z-
dc.date.available2020-07-08T03:19:08Z-
dc.date.issued2019-
dc.identifier.citationChin, 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.2019.04.020en_US
dc.identifier.issn1742-7061en_US
dc.identifier.urihttps://hdl.handle.net/10356/142907-
dc.description.abstractGenome 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.isoenen_US
dc.relation.ispartofActa Biomaterialiaen_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.subjectEngineering::Bioengineeringen_US
dc.titleScaffold-mediated non-viral delivery platform for CRISPR/Cas9-based genome editingen_US
dc.typeJournal Articleen
dc.contributor.schoolSchool of Chemical and Biomedical Engineeringen_US
dc.contributor.schoolInterdisciplinary Graduate School (IGS)en_US
dc.contributor.schoolLee Kong Chian School of Medicine (LKCMedicine)en_US
dc.contributor.organizationNTU Institute of Health Technologiesen_US
dc.identifier.doi10.1016/j.actbio.2019.04.020-
dc.description.versionAccepted versionen_US
dc.identifier.volume90en_US
dc.identifier.spage60en_US
dc.identifier.epage70en_US
dc.subject.keywordsGene Deliveryen_US
dc.subject.keywordsCas9 Proteinen_US
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