Type
ArticleKAUST Department
Bioscience ProgramBiological and Environmental Sciences and Engineering (BESE) Division
Plant Science
Desert Agriculture Initiative
Date
2019-12-02Online Publication Date
2019-12-02Print Publication Date
2019-12Permanent link to this record
http://hdl.handle.net/10754/660537
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BACKGROUND:CRISPR-Cas systems endow bacterial and archaeal species with adaptive immunity mechanisms to fend off invading phages and foreign genetic elements. CRISPR-Cas9 has been harnessed to confer virus interference against DNA viruses in eukaryotes, including plants. In addition, CRISPR-Cas13 systems have been used to target RNA viruses and the transcriptome in mammalian and plant cells. Recently, CRISPR-Cas13a has been shown to confer modest interference against RNA viruses. Here, we characterized a set of different Cas13 variants to identify those with the most efficient, robust, and specific interference activities against RNA viruses in planta using Nicotiana benthamiana. RESULTS:Our data show that LwaCas13a, PspCas13b, and CasRx variants mediate high interference activities against RNA viruses in transient assays. Moreover, CasRx mediated robust interference in both transient and stable overexpression assays when compared to the other variants tested. CasRx targets either one virus alone or two RNA viruses simultaneously, with robust interference efficiencies. In addition, CasRx exhibits strong specificity against the target virus and does not exhibit collateral activity in planta. CONCLUSIONS:Our data establish CasRx as the most robust Cas13 variant for RNA virus interference applications in planta and demonstrate its suitability for studying key questions relating to virus biology.Citation
Mahas, A., Aman, R., & Mahfouz, M. (2019). CRISPR-Cas13d mediates robust RNA virus interference in plants. Genome Biology, 20(1). doi:10.1186/s13059-019-1881-2Sponsors
We wish to thank members of the Laboratory for Genome Engineering and Synthetic Biology at King Abdullah University of Science and Technology for helpful discussions and comments on the manuscript.Publisher
Springer Science and Business Media LLCJournal
Genome biologyAdditional Links
https://genomebiology.biomedcentral.com/articles/10.1186/s13059-019-1881-2https://genomebiology.biomedcentral.com/track/pdf/10.1186/s13059-019-1881-2
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. DOI: 10.6084/m9.figshare.c.4766735 HANDLE: 10754/665165
ae974a485f413a2113503eed53cd6c53
10.1186/s13059-019-1881-2
Scopus Count
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