Please use this identifier to cite or link to this item:
https://hdl.handle.net/10356/87501
Title: | Systematic evaluation of CRISPR-Cas systems reveals design principles for genome editing in human cells | Authors: | Wang, Yuanming Liu, Ivy Kaiwen Sutrisnoh, Norfala-Aliah Srinivasan, Harini Zhang, Junyi Li, Jia Zhang, Fan Lalith, Charles Richard John Xing, Heyun Shanmugam, Raghuvaran Foo, Jia Nee Yeo, Hwee Ting Ooi, Kean Hean Bleckwehl, Tore Par, Rachel Yi Yun Lee, Shi Mun Ismail, Nur Nadiah Sanwari, Nur Aidah Lee, Vanessa Si Ting Lew, Jan Tan, Meng How |
Keywords: | CRISPR Genome Editing |
Issue Date: | 2018 | Source: | Wang, Y., Liu, I. K., Sutrisnoh, N.-A., Srinivasan, H., Zhang, J., Li, J., et al. (2018). Systematic evaluation of CRISPR-Cas systems reveals design principles for genome editing in human cells. Genome Biology, 19(1), 62-. | Series/Report no.: | Genome Biology | Abstract: | Background: While CRISPR-Cas systems hold tremendous potential for engineering the human genome, it is unclear how well each system performs against one another in both non-homologous end joining (NHEJ)-mediated and homology-directed repair (HDR)-mediated genome editing. Results: We systematically compare five different CRISPR-Cas systems in human cells by targeting 90 sites in genes with varying expression levels. For a fair comparison, we select sites that are either perfectly matched or have overlapping seed regions for Cas9 and Cpf1. Besides observing a trade-off between cleavage efficiency and target specificity for these natural endonucleases, we find that the editing activities of the smaller Cas9 enzymes from Staphylococcus aureus (SaCas9) and Neisseria meningitidis (NmCas9) are less affected by gene expression than the other larger Cas proteins. Notably, the Cpf1 nucleases from Acidaminococcus sp. BV3L6 and Lachnospiraceae bacterium ND2006 (AsCpf1 and LbCpf1, respectively) are able to perform precise gene targeting efficiently across multiple genomic loci using single-stranded oligodeoxynucleotide (ssODN) donor templates with homology arms as short as 17 nucleotides. Strikingly, the two Cpf1 nucleases exhibit a preference for ssODNs of the non-target strand sequence, while the popular Cas9 enzyme from Streptococcus pyogenes (SpCas9) exhibits a preference for ssODNs of the target strand sequence instead. Additionally, we find that the HDR efficiencies of Cpf1 and SpCas9 can be further improved by using asymmetric donors with longer arms 5′ of the desired DNA changes. Conclusions: Our work delineates design parameters for each CRISPR-Cas system and will serve as a useful reference for future genome engineering studies. | URI: | https://hdl.handle.net/10356/87501 http://hdl.handle.net/10220/45417 |
ISSN: | 1474-760X | DOI: | 10.1186/s13059-018-1445-x | Schools: | School of Chemical and Biomedical Engineering School of Biological Sciences Lee Kong Chian School of Medicine (LKCMedicine) |
Rights: | © 2018 The Author(s). This article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated. | Fulltext Permission: | open | Fulltext Availability: | With Fulltext |
Appears in Collections: | SCBE Journal Articles |
Files in This Item:
File | Description | Size | Format | |
---|---|---|---|---|
Systematic evaluation of CRISPR-Cas systems reveals design principles for genome editing in human cells.pdf | 2.11 MB | Adobe PDF | View/Open |
SCOPUSTM
Citations
10
54
Updated on Mar 28, 2024
Web of ScienceTM
Citations
5
117
Updated on Oct 26, 2023
Page view(s) 50
594
Updated on Mar 28, 2024
Download(s) 50
151
Updated on Mar 28, 2024
Google ScholarTM
Check
Altmetric
Items in DR-NTU are protected by copyright, with all rights reserved, unless otherwise indicated.