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dc.contributor.authorChooi, Wai Honen_US
dc.contributor.authorChin, Jiah Shinen_US
dc.contributor.authorChew, Sing Yianen_US
dc.identifier.citationChooi, W. H., Chin, J. S., & Chew, S. Y. (2021). Scaffold-based delivery of CRISPR/Cas9 ribonucleoproteins for genome editing. Methods in Molecular Biology, 2211, 183-191. doi:10.1007/978-1-0716-0943-9_13en_US
dc.description.abstractThe simple and versatile CRISPR/Cas9 system is a promising strategy for genome editing in mammalian cells. Generally, the genome editing components, namely Cas9 protein and single-guide RNA (sgRNA), are delivered in the format of plasmids, mRNA or ribonucleoprotein (RNP) complexes. In particular, non-viral approaches are desirable as they overcome the safety concerns posed by viral vectors. To control cell fate for tissue regeneration, scaffold-based delivery of genome editing components will offer a route for local delivery and provide possible synergistic effects with other factors such as topographical cues that are co-delivered by the same scaffold. In this chapter, we detail a simple method of surface modification to functionalize electrospun nanofibers with CRISPR/Cas9 RNP complexes. The mussel-inspired bio-adhesive coating will be used as it is a simple and effective method to immobilize biomolecules on the surface. Nanofibers will provide a biomimicking microenvironment and topographical cues to seeded cells. For evaluation, a model cell line with single copies of enhanced green fluorescent protein (U2OS.EGFP) will be used to validate the efficiency of gene disruption.en_US
dc.description.sponsorshipAgency for Science, Technology and Research (A*STAR)en_US
dc.description.sponsorshipMinistry of Education (MOE)en_US
dc.description.sponsorshipNational Medical Research Council (NMRC)en_US
dc.relation.ispartofMethods in Molecular Biologyen_US
dc.rights© 2021 Humana Press (Published by Springer). All rights reserved. This paper was published in Methods in Molecular Biology and is made available with permission of Humana Press (Published by Springer).en_US
dc.subjectEngineering::Chemical engineeringen_US
dc.titleScaffold-based delivery of CRISPR/Cas9 ribonucleoproteins for 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.description.versionAccepted versionen_US
dc.subject.keywordsGene deliveryen_US
dc.description.acknowledgementPartial funding support from the Singapore National Research Foundation under its National Medical Research Council-Cooperative Basic Research Grant (NMRC-CBRG) grant (NMRC/CBRG/0096/2015) and administered by the Singapore Ministry of Health’s National Medical Research Council; Ministry of Education Tier 1 grant (RG38/19); and A*Star BMRC Singapore-China 12th Joint Research Programme Grant (Project No: 1610500024) are acknowledged. Jiah Shin Chin would like to thank the NTU Interdisciplinary Graduate Research Officer’s scheme for supporting her through this work. We would also like to thank New England Biolabs for providing the Cas9 proteins.en_US
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