Multiplexed CRISPR technologies for gene editing and transcriptional regulation
Abstract
Multiplexed CRISPR technologies, in which numerous gRNAs or Cas enzymes are expressed at once, have facilitated powerful biological engineering applications, vastly enhancing the scope and efficiencies of genetic editing and transcriptional regulation. In this review, we discuss multiplexed CRISPR technologies and describe methods for the assembly, expression and processing of synthetic guide RNA arrays in vivo. Applications that benefit from multiplexed CRISPR technologies, including cellular recorders, genetic circuits, biosensors, combinatorial genetic perturbations, large-scale genome engineering and the rewiring of metabolic pathways, are highlighted. We also offer a glimpse of emerging challenges and emphasize experimental considerations for future studies.
Additional Information
© The Author(s) 2020. This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as 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 images or other third party material in this article are included in the article's Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/. Received: 27 October 2019; Accepted: 17 February 2020; Published online: 9 March 2020. R.L.A. received funding from Imperial College London in the form of an IC Research Fellowship and a BBSRC grant (BB/R01602X/1). These authors contributed equally: Alicia E. Graham, Lucie Studena. Author Contributions: R.L.-A. and N.S.M. conceived the paper. N.S.M. wrote most of the initial text with the help of L.S. and A.E.G. who contributed to the application and computational methods sections, respectively. R.L.-A provided guidance on the writing and direction of the review. All authors revised and finalized the paper. The authors declare no competing interests.Attached Files
Published - s41467-020-15053-x.pdf
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Additional details
- PMCID
- PMC7062760
- Eprint ID
- 101934
- Resolver ID
- CaltechAUTHORS:20200316-150529286
- BB/R01602X/1
- Biotechnology and Biological Sciences Research Council (BBSRC)
- Created
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2020-03-17Created from EPrint's datestamp field
- Updated
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2022-02-15Created from EPrint's last_modified field