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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.
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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