Pea early-browning virus -mediated genome editing via the CRISPR/Cas9 system in Nicotiana benthamiana and Arabidopsis
Name:
Pea early-browning-Manuscript.pdf
Size:
619.7Kb
Format:
PDF
Description:
Accepted Manuscript
Type
Article
KAUST Department
Biological and Environmental Sciences and Engineering (BESE) DivisionBioscience Program
Desert Agriculture Initiative
Laboratory for Genome Engineering
Plant Science
Plant Science Program
Date
2017-10-17Online Publication Date
2017-10-17Print Publication Date
2018-01Permanent link to this record
http://hdl.handle.net/10754/626046Metadata
Show full item recordAbstract
The clustered regularly interspaced palindromic repeats (CRISPR)/CRISPR-associated (Cas9) system has enabled efficient genome engineering in diverse plant species. However, delivery of genome engineering reagents, such as the single guide RNA (sgRNA), into plant cells remains challenging. Here, we report the engineering of Tobacco rattle virus (TRV) and Pea early browning virus (PEBV) to deliver one or multiple sgRNAs into Nicotiana benthamiana and Arabidopsis thaliana (Col-0) plants that overexpress a nuclear localization signal containing Cas9. Our data showed that TRV and PEBV can deliver sgRNAs into inoculated and systemic leaves, and this resulted in mutagenesis of the targeted genomic loci. Moreover, in N. benthamiana, PEBV-based sgRNA delivery resulted in more targeted mutations than TRV-based delivery. Our data indicate that TRV and PEBV can facilitate plant genome engineering and can be used to produce targeted mutations for functional analysis and other biotechnological applications across diverse plant species.Key message: Delivery of genome engineering reagents into plant cells is challenging and inefficient and this limit the applications of this technology in many plant species. RNA viruses such as TRV and PEBV provide an efficient tool to systemically deliver sgRNAs for targeted genome modification.Citation
Ali Z, Eid A, Ali S, Mahfouz MM (2017) Pea early-browning virus -mediated genome editing via the CRISPR/Cas9 system in Nicotiana benthamiana and Arabidopsis. Virus Research. Available: http://dx.doi.org/10.1016/j.virusres.2017.10.009.Sponsors
We would like to thank member of the laboratory for genome engineering for continuous discussions. We would like to thank Professor Elisabeth Johansen, Danish Institute for food and veterinary research, for providing PEBV-containing binary constructs. This study is supported by King Abdullah University of Science and Technology (KAUST).Publisher
Elsevier BVJournal
Virus ResearchPubMed ID
29051052Additional Links
http://www.sciencedirect.com/science/article/pii/S0168170217305543Scopus Count
Related items
Showing items related by title, author, creator and subject.
-
Phylogenetic analysis of Melon chlorotic leaf curl virus from Guatemala: Another emergent species in the Squash leaf curl virus clade(Virus Research, Elsevier BV, 2011-06) [Article]The genome of a new bipartite begomovirus Melon chlorotic leaf curl virus from Guatemala (MCLCuV-GT) was cloned and the genome sequence was determined. The virus causes distinct symptoms on melons that were not previously observed in melon crops in Guatemala or elsewhere. Phylogenetic analysis of MCLCuV-GT and begomoviruses infecting cucurbits and other host plant species indicated that its closest relative was MCLCuV from Costa Rica (MCLCuV-CR). The DNA-A components of two isolates shared 88.8% nucleotide identity, making them strains of the same species. Further, both MCLCuV-GT and MCLCuV-CR grouped with other Western Hemisphere cucurbit-infecting species in the SLCV-clade making them the most southerly cucurbit-infecting members of the clade to date. Although the common region of the cognate components of MCLCuV-GT and MCLCuV-CR, shared similar to 96.3% nucleotide identity. While DNA-A and DNA-B components of MCLCuV-GT were less than 86% nucleotide identity with the respective DNAA and DNA-B common regions of MCLCuV-CR. The late viral genes of the two strains shared the least nt identity (<88%) while their early genes shared the highest nt identity (>90%). The collective evidence suggests that these two strains of MCLCuV are evolutionarily divergent owing in part to recombination, but also due to the accumulation of a substantial number of mutations. In addition they are differentially host-adapted, as has been documented for other cucurbit-infecting, bean-adapted, species in the SLCV clade. (C) 2011 Elsevier B.V. All rights reserved.
-
Viruses-to-mobile genetic elements skew in the deep Atlantis II brine pool sediments(Scientific Reports, Springer Nature, 2016-09-06) [Article]The central rift of the Red Sea has 25 brine pools with different physical and geochemical characteristics. Atlantis II (ATIID), Discovery Deeps (DD) and Chain Deep (CD) are characterized by high salinity, temperature and metal content. Several studies reported microbial communities in these brine pools, but few studies addressed the brine pool sediments. Therefore, sediment cores were collected from ATIID, DD, CD brine pools and an adjacent brine-influenced site. Sixteen different lithologic sediment sections were subjected to shotgun DNA pyrosequencing to generate 1.47 billion base pairs (1.47 × 109 bp). We generated sediment-specific reads and attempted to annotate all reads. We report the phylogenetic and biochemical uniqueness of the deepest ATIID sulfur-rich brine pool sediments. In contrary to all other sediment sections, bacteria dominate the deepest ATIID sulfur-rich brine pool sediments. This decrease in virus-to-bacteria ratio in selected sections and depth coincided with an overrepresentation of mobile genetic elements. Skewing in the composition of viruses-to-mobile genetic elements may uniquely contribute to the distinct microbial consortium in sediments in proximity to hydrothermally active vents of the Red Sea and possibly in their surroundings, through differential horizontal gene transfer.
-
System Modeling of Virus Transmission and Detection in Molecular Communication Channels(2018 IEEE International Conference on Communications (ICC), Institute of Electrical and Electronics Engineers (IEEE), 2018-08-20) [Conference Paper]Aerosol Transmission is one of the major spread mechanism for diseases and is responsible for transmission of virus over long distances. The advancement in nanotechnology has resulted in sensors and systems that allow us to deal with nanosized biological entities such as virus and bacteria. In this work, the idea of viewing virus transmission through aerosols and their transport as a molecular communication problem is introduced. In such problems one has little or no control over transmission, however, a robust receiver can be designed using nano-biosensors for information extraction. Thus, the objective of this work is to treat viral aerosol spread as a blind communication problem and present a mathematical model for it. Specifically, we study the virus transmission from an engineering perspective and derive an end-to-end mathematical model for virus transmission in the atmosphere. The receiver architecture composed of air sampler and Silicon Nanowire field effect transistor is also discussed. Furthermore, a detection problem is formulated and simulation results are reported that justify the feasibility of such setups in bio-monitoring applications.