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Title: Scaffold-based delivery of CRISPR/Cas9 ribonucleoproteins for genome editing
Authors: Chooi, Wai Hon
Chin, Jiah Shin
Chew, Sing Yian
Keywords: Engineering::Chemical engineering
Issue Date: 2020
Source: Chooi, 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_13
Journal: Methods in Molecular Biology
Abstract: The 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.
ISSN: 1573-4978
DOI: 10.1007/978-1-0716-0943-9_13
Schools: School of Chemical and Biomedical Engineering 
Interdisciplinary Graduate School (IGS) 
Lee Kong Chian School of Medicine (LKCMedicine) 
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).
Fulltext Permission: open
Fulltext Availability: With Fulltext
Appears in Collections:SCBE Journal Articles

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