Scaffold-based delivery of CRISPR/Cas9 ribonucleoproteins for genome editing
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, no...
| Main Authors: | , , |
|---|---|
| Other Authors: | |
| Format: | Journal Article |
| Language: | English |
| Published: |
2021
|
| Subjects: | |
| Online Access: | https://hdl.handle.net/10356/145891 |
| _version_ | 1811687483257126912 |
|---|---|
| author | Chooi, Wai Hon Chin, Jiah Shin Chew, Sing Yian |
| author2 | School of Chemical and Biomedical Engineering |
| author_facet | School of Chemical and Biomedical Engineering Chooi, Wai Hon Chin, Jiah Shin Chew, Sing Yian |
| author_sort | Chooi, Wai Hon |
| collection | NTU |
| description | 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. |
| first_indexed | 2024-10-01T05:17:02Z |
| format | Journal Article |
| id | ntu-10356/145891 |
| institution | Nanyang Technological University |
| language | English |
| last_indexed | 2024-10-01T05:17:02Z |
| publishDate | 2021 |
| record_format | dspace |
| spelling | ntu-10356/1458912023-12-29T06:49:01Z Scaffold-based delivery of CRISPR/Cas9 ribonucleoproteins for genome editing Chooi, Wai Hon Chin, Jiah Shin Chew, Sing Yian School of Chemical and Biomedical Engineering Interdisciplinary Graduate School (IGS) Lee Kong Chian School of Medicine (LKCMedicine) Engineering::Chemical engineering Gene delivery Electrospinning 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. Agency for Science, Technology and Research (A*STAR) Ministry of Education (MOE) National Medical Research Council (NMRC) Accepted version Partial 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. 2021-01-13T07:40:29Z 2021-01-13T07:40:29Z 2020 Journal Article 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 1573-4978 https://hdl.handle.net/10356/145891 10.1007/978-1-0716-0943-9_13 2211 183 191 en Methods in Molecular Biology © 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). application/pdf |
| spellingShingle | Engineering::Chemical engineering Gene delivery Electrospinning Chooi, Wai Hon Chin, Jiah Shin Chew, Sing Yian Scaffold-based delivery of CRISPR/Cas9 ribonucleoproteins for genome editing |
| title | Scaffold-based delivery of CRISPR/Cas9 ribonucleoproteins for genome editing |
| title_full | Scaffold-based delivery of CRISPR/Cas9 ribonucleoproteins for genome editing |
| title_fullStr | Scaffold-based delivery of CRISPR/Cas9 ribonucleoproteins for genome editing |
| title_full_unstemmed | Scaffold-based delivery of CRISPR/Cas9 ribonucleoproteins for genome editing |
| title_short | Scaffold-based delivery of CRISPR/Cas9 ribonucleoproteins for genome editing |
| title_sort | scaffold based delivery of crispr cas9 ribonucleoproteins for genome editing |
| topic | Engineering::Chemical engineering Gene delivery Electrospinning |
| url | https://hdl.handle.net/10356/145891 |
| work_keys_str_mv | AT chooiwaihon scaffoldbaseddeliveryofcrisprcas9ribonucleoproteinsforgenomeediting AT chinjiahshin scaffoldbaseddeliveryofcrisprcas9ribonucleoproteinsforgenomeediting AT chewsingyian scaffoldbaseddeliveryofcrisprcas9ribonucleoproteinsforgenomeediting |