Multiplexed heritable gene editing using RNA viruses and mobile single guide RNAs

Evan E Ellison^{1

2

3

4}, Ugrappa Nagalakshmi⁵, Maria Elena Gamo^{1

2

3}, Pin-Jui Huang⁵, Savithramma Dinesh-Kumar⁵, Daniel F Voytas^{6

7

8}

Affiliations

¹ Department of Genetics, Cell Biology and Development, University of Minnesota, St. Paul, MN, USA.
² Center for Precision Plant Genomics, University of Minnesota, St. Paul, MN, USA.
³ Center for Genome Engineering, University of Minnesota, St. Paul, MN, USA.
⁴ Plant and Microbial Biology Graduate Program, University of Minnesota, St. Paul, MN, USA.
⁵ Department of Plant Biology and The Genome Center, College of Biological Sciences, University of California, Davis, Davis, CA, USA.
⁶ Department of Genetics, Cell Biology and Development, University of Minnesota, St. Paul, MN, USA. voytas@umn.edu.
⁷ Center for Precision Plant Genomics, University of Minnesota, St. Paul, MN, USA. voytas@umn.edu.
⁸ Center for Genome Engineering, University of Minnesota, St. Paul, MN, USA. voytas@umn.edu.

PMID: 32483329
DOI: 10.1038/s41477-020-0670-y

Multiplexed heritable gene editing using RNA viruses and mobile single guide RNAs

Evan E Ellison et al. Nat Plants. 2020 Jun.

. 2020 Jun;6(6):620-624.

doi: 10.1038/s41477-020-0670-y. Epub 2020 Jun 1.

Authors

Evan E Ellison^{1

2

3

4}, Ugrappa Nagalakshmi⁵, Maria Elena Gamo^{1

2

3}, Pin-Jui Huang⁵, Savithramma Dinesh-Kumar⁵, Daniel F Voytas^{6

7

8}

Affiliations

¹ Department of Genetics, Cell Biology and Development, University of Minnesota, St. Paul, MN, USA.
² Center for Precision Plant Genomics, University of Minnesota, St. Paul, MN, USA.
³ Center for Genome Engineering, University of Minnesota, St. Paul, MN, USA.
⁴ Plant and Microbial Biology Graduate Program, University of Minnesota, St. Paul, MN, USA.
⁵ Department of Plant Biology and The Genome Center, College of Biological Sciences, University of California, Davis, Davis, CA, USA.
⁶ Department of Genetics, Cell Biology and Development, University of Minnesota, St. Paul, MN, USA. voytas@umn.edu.
⁷ Center for Precision Plant Genomics, University of Minnesota, St. Paul, MN, USA. voytas@umn.edu.
⁸ Center for Genome Engineering, University of Minnesota, St. Paul, MN, USA. voytas@umn.edu.

PMID: 32483329
DOI: 10.1038/s41477-020-0670-y

Erratum in

Author Correction: Multiplexed heritable gene editing using RNA viruses and mobile single guide RNAs.
Ellison EE, Nagalakshmi U, Gamo ME, Huang PJ, Dinesh-Kumar S, Voytas DF. Ellison EE, et al. Nat Plants. 2021 Jan;7(1):99. doi: 10.1038/s41477-020-00837-2. Nat Plants. 2021. PMID: 33328598 No abstract available.

Abstract

An in planta gene editing approach was developed wherein Cas9 transgenic plants are infected with an RNA virus that expresses single guide RNAs (sgRNAs). The sgRNAs are augmented with sequences that promote cell-to-cell mobility. Mutant progeny are recovered in the next generation at frequencies ranging from 65 to 100%; up to 30% of progeny derived from plants infected with a virus expressing three sgRNAs have mutations in all three targeted loci.

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Comment in

The way to true plant genome editing.
Kim SG. Kim SG. Nat Plants. 2020 Jul;6(7):736-737. doi: 10.1038/s41477-020-0723-2. Nat Plants. 2020. PMID: 32665647 No abstract available.

References

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1. Altpeter, F. et al. Advancing crop transformation in the era of genome editing. Plant Cell 28, 1510–1520 (2016). - PubMed - PMC
1. Clough, S. J. & Bent, A. F. Floral dip: a simplified method for Agrobacterium-mediated transformation of Arabidopsis thaliana. Plant J. 16, 735–743 (1998). - DOI
1. Hamada, H. et al. An in planta biolistic method for stable wheat transformation. Sci. Rep. 7, 11443 (2017). - DOI
1. Burch-Smith, T. M. et al. Applications and advantages of virus-induced gene silencing for gene function studies in plants. Plant J. 39, 734–746 (2004). - DOI

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Multiplexed heritable gene editing using RNA viruses and mobile single guide RNAs

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Multiplexed heritable gene editing using RNA viruses and mobile single guide RNAs

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