Journal
PLOS ONE
Volume 11, Issue 9, Pages -Publisher
PUBLIC LIBRARY SCIENCE
DOI: 10.1371/journal.pone.0162169
Keywords
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Categories
Funding
- University of North Carolina at Chapel Hill
- Virginia Tech
- Pew Latin American Fellows Program in the Biomedical Sciences [00026198]
- National Science Foundation-Division of Integrated Organismal Systems [IOS-1455607]
- Howard Hughes Medical Institute - HHMI
- Gordon and Betty Moore Foundation [GBMF3030]
- UNC
- Howard HughesMedical Institute
- Gordon and BettyMoore Foundation [GBMF3030]
- NSF-IOS [IOS-1455607]
- HHMI
- Direct For Biological Sciences
- Division Of Integrative Organismal Systems [1257373] Funding Source: National Science Foundation
- Division Of Integrative Organismal Systems
- Direct For Biological Sciences [1455607] Funding Source: National Science Foundation
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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 thatmultiplex 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 cleanmutant generation with no need to enhance specificity or adopt alternate Cas9 variants.
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