Genome-Wide Assessment of Efficiency and Specificity in CRISPR/Cas9 Mediated Multiple Site Targeting in Arabidopsis
Public Deposited- Creator
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Peterson, Brenda A.
- Affiliation: College of Arts and Sciences, Department of Biology
- Other Affiliation: Departmentof Biological Sciences; Virginia Tech
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Haak, David C.
- Other Affiliation: Department of Plant Pathology Physiology; andWeed Science; Virginia Tech
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Nishimura, Marc T.
- Affiliation: University of North Carolina at Chapel Hill
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Teixeira, Paulo J. P. L.
- Affiliation: University of North Carolina at Chapel Hill
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James, Sean R.
- Affiliation: College of Arts and Sciences, Department of Biology
- Other Affiliation: Departmentof Biological Sciences; Virginia Tech
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Dangl, Jeffery L.
- Affiliation: College of Arts and Sciences, Department of Biology
- Other Affiliation: University of North Carolina at Chapel Hill
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Nimchuk, Zachary L.
- Affiliation: College of Arts and Sciences, Department of Biology
- Other Affiliation: Departmentof Biological Sciences; Virginia Tech
- Abstract
- Simultaneous multiplex mutation of large gene families using Cas9 has the potential to revolutionize agriculture and plant sciences. The targeting of multiple genomic sites at once raises concerns about the efficiency and specificity in targeting. The model Arabidopsis thaliana is widely used in basic plant research. Previous work has suggested that the Cas9 off-target rate in Arabidopsis is undetectable. Here we use deep sequencing on pooled plants simultaneously targeting 14 distinct genomic loci to demonstrate that multiplex targeting in Arabidopsis is highly specific to on-target sites with no detectable off-target events. In addition, chromosomal translocations are extremely rare. The high specificity of Cas9 in Arabidopsis makes this a reliable method for clean mutant generation with no need to enhance specificity or adopt alternate Cas9 variants.
- Date of publication
- 2016
- Keyword
- DOI
- Identifier
- PMID: 27622539
- Onescience id: 07d5e9a26786e61fb98cdd925bcd0d8ca764eb60
- Publisher DOI: https://doi.org/10.1371/journal.pone.0162169
- PMCID: PMC5021288
- Resource type
- Article
- Rights statement
- In Copyright
- Journal title
- PloS One
- Journal volume
- 11
- Journal issue
- 9
- Page start
- e0162169
- Language
- English
- ISSN
- 1932-6203
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