Protein expression and gene editing in monocots using foxtail mosaic virus vectors

Mei, Yu; Beernink, Bliss M.; Ellison, Evan E.; Konečná, Eva; Neelakandan, Anjanasree K.; Voytas, Daniel F.; Whitham, Steven A.

doi:10.1002/pld3.181

Title: Protein expression and gene editing in monocots using foxtail mosaic virus vectors

Journal Article · Fri Nov 22 00:00:00 EST 2019 · Plant Direct

DOI:https://doi.org/10.1002/pld3.181· OSTI ID:1670835

Mei, Yu ^[1]; Beernink, Bliss M. ^[1]; Ellison, Evan E. ^[2]; Konečná, Eva ^[2]; Neelakandan, Anjanasree K. ^[3]; Voytas, Daniel F. ^[2]; Whitham, Steven A. ^[1]

Department of Plant Pathology and Microbiology Iowa State University Ames IA USA
Department of Genetics, Cell Biology and Development Center for Genome Engineering Center for Precision Plant Genomics University of Minnesota St. Paul MN USA
Department of Genetics, Development and Cell Biology Iowa State University Ames IA USA

Plant viruses can be engineered to carry sequences that direct silencing of target host genes, expression of heterologous proteins, or editing of host genes. A set of foxtail mosaic virus (FoMV) vectors was developed that can be used for transient gene expression and single guide RNA delivery for Cas9-mediated gene editing in maize, Setaria viridis, and Nicotiana benthamiana. This was accomplished by duplicating the FoMV capsid protein subgenomic promoter, abolishing the unnecessary open reading frame 5A, and inserting a cloning site containing unique restriction endonuclease cleavage sites immediately after the duplicated promoter. The modified FoMV vectors transiently expressed green fluorescent protein (GFP) and bialaphos resistance (BAR) protein in leaves of systemically infected maize seedlings. GFP was detected in epidermal and mesophyll cells by epifluorescence microscopy, and expression was confirmed by Western blot analyses. Plants infected with FoMV carrying the bar gene were temporarily protected from a glufosinate herbicide, and expression was confirmed using a rapid antibody-based BAR strip test. Expression of these proteins was stabilized by nucleotide substitutions in the sequence of the duplicated promoter region. Single guide RNAs expressed from the duplicated promoter mediated edits in the N. benthamiana Phytoene desaturase gene, the S. viridis Carbonic anhydrase 2 gene, and the maize HKT1 gene encoding a potassium transporter. The efficiency of editing was enhanced in the presence of synergistic viruses and a viral silencing suppressor. This work expands the utility of FoMV for virus-induced gene silencing (VIGS), virus-mediated overexpression (VOX), and virus-enabled gene editing (VEdGE) in monocots.

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Research Organization:: Donald Danforth Plant Science Center, St. Louis, MO (United States)

Sponsoring Organization:: USDOE Office of Science (SC)

Grant/Contract Number:: DE‐SC0018277; SC0018277

OSTI ID:: 1670835

Alternate ID(s):: OSTI ID: 1670840; OSTI ID: 1803339; OSTI ID: 2308797

Journal Information:: Plant Direct, Journal Name: Plant Direct Vol. 3 Journal Issue: 11; ISSN 2475-4455

Publisher:: Wiley Blackwell (John Wiley & Sons)Copyright Statement

Country of Publication:: United Kingdom

Language:: English

References (48)

Harnessed viruses in the age of metagenomics and synthetic biology: an update on infectious clone assembly and biotechnologies of plant viruses Pasin, Fabio; Menzel, Wulf; Daròs, José‐Antonio Plant Biotechnology Journal, Vol. 17, Issue 6 https://doi.org/10.1111/pbi.13084	journal	February 2019
Development of a Virus-Induced Gene-Silencing System for Hexaploid Wheat and Its Use in Functional Analysis of the Lr21 -Mediated Leaf Rust Resistance Pathway Scofield, Steven R.; Huang, Li; Brandt, Amanda S. Plant Physiology, Vol. 138, Issue 4 https://doi.org/10.1104/pp.105.061861	journal	July 2005
Virus-induced gene silencing in rice using a vector derived from a DNA virus Purkayastha, Arunima; Mathur, Saloni; Verma, Vidhu Planta, Vol. 232, Issue 6 https://doi.org/10.1007/s00425-010-1273-z	journal	September 2010
An efficient Foxtail mosaic virus vector system with reduced environmental risk Liu, Zun; Kearney, Christopher M. BMC Biotechnology, Vol. 10, Issue 1 https://doi.org/10.1186/1472-6750-10-88	journal	December 2010
Use of Potato Virus X (PVX)–Based Vectors for Gene Expression and Virus‐Induced Gene Silencing (VIGS) Lacomme, Christophe; Chapman, Sean Current Protocols in Microbiology, Vol. 8, Issue 1 https://doi.org/10.1002/9780471729259.mc16i01s8	journal	February 2008
An efficient virus-induced gene silencing vector for maize functional genomics research Wang, Rong; Yang, Xinxin; Wang, Nian The Plant Journal, Vol. 86, Issue 1 https://doi.org/10.1111/tpj.13142	journal	April 2016
Viral Vectors for Plant Genome Engineering Zaidi, Syed Shan-e-Ali; Mansoor, Shahid Frontiers in Plant Science, Vol. 8 https://doi.org/10.3389/fpls.2017.00539	journal	April 2017
Plant viral synergism: the potyviral genome encodes a broad-range pathogenicity enhancer that transactivates replication of heterologous viruses. Pruss, G.; Ge, X.; Shi, X. M. The Plant Cell, Vol. 9, Issue 6 https://doi.org/10.1105/tpc.9.6.859	journal	June 1997
Virus-Induced Gene Silencing in Maize with a Foxtail mosaic virus Vector Mei, Yu; Whitham, Steven A. Maize https://doi.org/10.1007/978-1-4939-7315-6_7	book	September 2017
Gene Targeting by Homology-Directed Repair in Rice Using a Geminivirus-Based CRISPR/Cas9 System Wang, Mugui; Lu, Yuming; Botella, José Ramón Molecular Plant, Vol. 10, Issue 7 https://doi.org/10.1016/j.molp.2017.03.002	journal	July 2017
Cell-to-Cell and Phloem-Mediated Transport of Potato Virus X: The Role of Virions Cruz, Simon Santa; Roberts, Alison G.; Prior, Denton A. M. The Plant Cell, Vol. 10, Issue 4 https://doi.org/10.1105/tpc.10.4.495	journal	April 1998
Barley Stripe Mosaic Virus - Mediated Tools for Investigating Gene Function in Cereal Plants and Their Pathogens: Virus-Induced Gene Silencing, Host-Mediated Gene Silencing, and Virus-Mediated Overexpression of Heterologous Protein Lee, Wing-Sham; Hammond-Kosack, Kim E.; Kanyuka, Kostya Plant Physiology, Vol. 160, Issue 2 https://doi.org/10.1104/pp.112.203489	journal	August 2012
Development of new potato virus X-based vectors for gene over-expression and gene silencing assay Wang, Ying; Cong, Qian-Qian; Lan, Yu-Fei Virus Research, Vol. 191 https://doi.org/10.1016/j.virusres.2014.07.018	journal	October 2014
Conferring resistance to geminiviruses with the CRISPR–Cas prokaryotic immune system Baltes, Nicholas J.; Hummel, Aaron W.; Konecna, Eva Nature Plants, Vol. 1, Issue 10 https://doi.org/10.1038/nplants.2015.145	journal	September 2015
Revised sequence of foxtail mosaic virus reveals a triple gene block structure similar to potato virus X Bruun-Rasmussen, M.; Madsen, C. T.; Johansen, E. Archives of Virology, Vol. 153, Issue 1 https://doi.org/10.1007/s00705-007-1057-3	journal	October 2007
Development of Beet necrotic yellow vein virus ‐based vectors for multiple‐gene expression and guide RNA delivery in plant genome editing Jiang, Ning; Zhang, Chao; Liu, Jun‐Ying Plant Biotechnology Journal, Vol. 17, Issue 7 https://doi.org/10.1111/pbi.13055	journal	January 2019
Rescue of a plant cytorhabdovirus as versatile expression platforms for planthopper and cereal genomic studies Gao, Qiang; Xu, Wen‐Ya; Yan, Teng New Phytologist, Vol. 223, Issue 4 https://doi.org/10.1111/nph.15889	journal	April 2019
Optimizing virus-induced gene silencing efficiency with Cymbidium mosaic virus in Phalaenopsis flower Hsieh, Ming-Hsien; Lu, Hsiang-Chia; Pan, Zhao-Jun Plant Science, Vol. 201-202 https://doi.org/10.1016/j.plantsci.2012.11.003	journal	March 2013
Co-expression of multiple target proteins in plants from a tobacco mosaic virus vector using a combination of homologous and heterologous subgenomic promoters Roy, Gourgopal; Weisburg, Sangeetha; Foy, Kelly Archives of Virology, Vol. 156, Issue 11 https://doi.org/10.1007/s00705-011-1069-x	journal	July 2011
The Development of an Efficient Multipurpose Bean Pod Mottle Virus Viral Vector Set for Foreign Gene Expression and RNA Silencing Zhang, Chunquan; Bradshaw, Jeffrey D.; Whitham, Steven A. Plant Physiology, Vol. 153, Issue 1 https://doi.org/10.1104/pp.109.151639	journal	March 2010
Foxtail Mosaic Virus -Induced Gene Silencing in Monocot Plants Liu, Na; Xie, Ke; Jia, Qi Plant Physiology, Vol. 171, Issue 3 https://doi.org/10.1104/pp.16.00010	journal	May 2016
The Complete Nucleotide Sequence of Potato Virus X and Its Homologies at the Amino Acid Level with Various Plus-stranded RNA Viruses Huisman, M. J.; Linthorst, H. J. M.; Bol, J. F. Journal of General Virology, Vol. 69, Issue 8 https://doi.org/10.1099/0022-1317-69-8-1789	journal	August 1988
Pea early-browning virus-mediated genome editing via the CRISPR/Cas9 system in Nicotiana benthamiana and Arabidopsis Ali, Zahir; Eid, Ayman; Ali, Shakila Virus Research, Vol. 244 https://doi.org/10.1016/j.virusres.2017.10.009	journal	January 2018
Contrasting effects of HC-Pro and 2b viral suppressors from Sugarcane mosaic virus and Tomato aspermy cucumovirus on the accumulation of siRNAs Zhang, Xiaoming; Du, Peng; Lu, Lu Virology, Vol. 374, Issue 2 https://doi.org/10.1016/j.virol.2007.12.045	journal	May 2008
Characterization of a Brome mosaic virus Strain and Its Use as a Vector for Gene Silencing in Monocotyledonous Hosts Ding, Xin Shun; Schneider, William L.; Chaluvadi, Srinivasa Rao Molecular Plant-Microbe Interactions, Vol. 19, Issue 11 https://doi.org/10.1094/MPMI-19-1229	journal	November 2006
Potato virus X -based expression vectors are stabilized for long-term production of proteins and larger inserts Dickmeis, Christina; Fischer, Rainer; Commandeur, Ulrich Biotechnology Journal, Vol. 9, Issue 11 https://doi.org/10.1002/biot.201400347	journal	October 2014
Efficient Virus-Mediated Genome Editing in Plants Using the CRISPR/Cas9 System Ali, Zahir; Abul-faraj, Aala; Li, Lixin Molecular Plant, Vol. 8, Issue 8 https://doi.org/10.1016/j.molp.2015.02.011	journal	August 2015
Multiplexed Gene Editing and Protein Overexpression Using a Tobacco mosaic virus Viral Vector Cody, Will B.; Scholthof, Herman B.; Mirkov, T. Erik Plant Physiology, Vol. 175, Issue 1 https://doi.org/10.1104/pp.17.00411	journal	June 2017
The open reading frame 5A of foxtail mosaic virus is expressed in vivo and is dispensable for systemic infection Robertson, N. L.; French, R.; Morris, T. J. Archives of Virology, Vol. 145, Issue 8 https://doi.org/10.1007/s007050070084	journal	August 2000
Expression of a flower-specific Myb protein in leaf cells using a viral vector causes ectopic activation of a target promoter. Sablowski, R. W.; Baulcombe, D. C.; Bevan, M. Proceedings of the National Academy of Sciences, Vol. 92, Issue 15 https://doi.org/10.1073/pnas.92.15.6901	journal	July 1995
High-efficiency gene targeting in hexaploid wheat using DNA replicons and CRISPR/Cas9 Gil-Humanes, Javier; Wang, Yanpeng; Liang, Zhen The Plant Journal, Vol. 89, Issue 6 https://doi.org/10.1111/tpj.13446	journal	February 2017
Mutations That Alter a Conserved Element Upstream of the Potato Virus X Triple Block and Coat Protein Genes Affect Subgenomic RNA Accumulation Kim, Kook-Hyung; Hemenway, Cynthia Virology, Vol. 232, Issue 1 https://doi.org/10.1006/viro.1997.8565	journal	May 1997
Mutations in the Region Encoding the Central Domain of Helper Component-Proteinase (HC-Pro) Eliminate Potato Virus X/Potyviral Synergism Shi, Xing Ming; Miller, Heather; Verchot, Jeanmarie Virology, Vol. 231, Issue 1 https://doi.org/10.1006/viro.1997.8488	journal	April 1997
Geminivirus-Mediated Genome Editing in Potato (Solanum tuberosum L.) Using Sequence-Specific Nucleases Butler, Nathaniel M.; Baltes, Nicholas J.; Voytas, Daniel F. Frontiers in Plant Science, Vol. 7 https://doi.org/10.3389/fpls.2016.01045	journal	July 2016
A plant virus vector for systemic expression of foreign genes in cereals Choi, Il-Ryong; Stenger, Drake C.; Morris, T. Jack The Plant Journal, Vol. 23, Issue 4 https://doi.org/10.1046/j.1365-313x.2000.00820.x	journal	August 2000
An Agrobacterium -delivered CRISPR/Cas9 system for high-frequency targeted mutagenesis in maize Char, Si Nian; Neelakandan, Anjanasree K.; Nahampun, Hartinio Plant Biotechnology Journal, Vol. 15, Issue 2 https://doi.org/10.1111/pbi.12611	journal	September 2016
Purification and Partial Characterization of Maize Dwarf Mosaic Virus Strain B (Sugarcane Mosaic Virus) Hill, J. H.; Ford, R. E.; Benner, H. I. Journal of General Virology, Vol. 20, Issue 3 https://doi.org/10.1099/0022-1317-20-3-327	journal	September 1973
A dual gene-silencing vector system for monocot and dicot plants Liou, Ming-Ru; Huang, Ying-Wen; Hu, Chung-Chi Plant Biotechnology Journal, Vol. 12, Issue 3 https://doi.org/10.1111/pbi.12140	journal	November 2013
Foxtail mosaic virus : A Viral Vector for Protein Expression in Cereals Bouton, Clément; King, Robert C.; Chen, Hongxin Plant Physiology, Vol. 177, Issue 4 https://doi.org/10.1104/pp.17.01679	journal	June 2018
Stability of Potato virus X expression vectors is related to insert size: implications for replication models and risk assessment Avesani, Linda; Marconi, Giampiero; Morandini, Francesca Transgenic Research, Vol. 16, Issue 5 https://doi.org/10.1007/s11248-006-9051-1	journal	January 2007
Purification and Properties of Foxtail Mosaic Virus Paulsen, A. Q. Phytopathology, Vol. 77, Issue 11 https://doi.org/10.1094/Phyto-67-1346	journal	January 1977
Optimized sgRNA design to maximize activity and minimize off-target effects of CRISPR-Cas9 Doench, John G.; Fusi, Nicolo; Sullender, Meagan Nature Biotechnology, Vol. 34, Issue 2 https://doi.org/10.1038/nbt.3437	journal	January 2016
A Multipurpose Toolkit to Enable Advanced Genome Engineering in Plants Čermák, Tomáš; Curtin, Shaun J.; Gil-Humanes, Javier The Plant Cell, Vol. 29, Issue 6 https://doi.org/10.1105/tpc.16.00922	journal	May 2017
Selectable viruses and altered susceptibility mutants in Arabidopsis thaliana Whitham, S. A.; Yamamoto, M. L.; Carrington, J. C. Proceedings of the National Academy of Sciences, Vol. 96, Issue 2 https://doi.org/10.1073/pnas.96.2.772	journal	January 1999
Manipulating plant RNA-silencing pathways to improve the gene editing efficiency of CRISPR/Cas9 systems Mao, Yanfei; Yang, Xiaoxuan; Zhou, Yiting Genome Biology, Vol. 19, Issue 1 https://doi.org/10.1186/s13059-018-1529-7	journal	September 2018
Efficient and stable expression of GFP through Wheat streak mosaic virus-based vectors in cereal hosts using a range of cleavage sites: Formation of dense fluorescent aggregates for sensitive virus tracking Tatineni, Satyanarayana; McMechan, Anthony J.; Hein, Gary L. Virology, Vol. 410, Issue 1 https://doi.org/10.1016/j.virol.2010.10.043	journal	February 2011
A CRISPR/Cas9 toolkit for multiplex genome editing in plants Xing, Hui-Li; Dong, Li; Wang, Zhi-Ping BMC Plant Biology, Vol. 14, Issue 1 https://doi.org/10.1186/s12870-014-0327-y	journal	November 2014
Efficient in planta gene targeting in tomato using geminiviral replicons and the CRISPR/Cas9 system Dahan-Meir, Tal; Filler-Hayut, Shdema; Melamed-Bessudo, Cathy The Plant Journal, Vol. 95, Issue 1 https://doi.org/10.1111/tpj.13932	journal	May 2018

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Title: Protein expression and gene editing in monocots using foxtail mosaic virus vectors

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