Journal
PLANT CELL REPORTS
Volume 36, Issue 6, Pages 1005-1008Publisher
SPRINGER
DOI: 10.1007/s00299-017-2125-0
Keywords
Genome editing; CRISPR/Cas9; Arabidopsis thaliana; Polyploidy; Genome dosage
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Funding
- Science Foundation Ireland (SFI) [13/IA/1820]
- Irish Research Council (IRC)
- Science Foundation Ireland (SFI) [13/IA/1820] Funding Source: Science Foundation Ireland (SFI)
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RNA-guided endonuclease-mediated targeted mutagenesis using the clustered regularly interspersed short palindromic repeats (CRISPR)/Cas9 system has been successful at targeting specific loci for modification in plants. While polyploidy is an evolutionary mechanism enabling plant adaptation, the analysis of gene function in polyploid plants has been limited due to challenges associated with generating polyploid knockout mutants for all gene copies in polyploid plant lines. This study investigated whether CRISPR/Cas9 mediated targeted mutagenesis can generate nulliplex tetraploid mutant lines in Arabidopsis thaliana, while also comparing the relative efficiency of targeted mutagenesis in tetraploid (4x) versus diploid (2x) backgrounds. Using CRISPR/Cas9 genome editing to generate knockout alleles of the TTG1 gene, we demonstrate that homozygous nulliplex mutants can be directly generated in tetraploid Arabidopsis thaliana plants. CRISPR/Cas9 genome editing now provides a route to more efficient generation of polyploid mutants for improving understanding of genome dosage effects in plants.
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