CRISPR-Cas9 Gene Editing of the Gene Family in Wheat.

The highly conserved encodes a bifunctional enzyme with inositol polyphosphate-1-phosphatase and 3' (2'), 5'-bisphosphate nucleotidase activity and has been shown to alter abiotic stress tolerance in plants when disrupted. Precise gene editing techniques were used to generate mutants in hexaploid bread wheat. The CRISPR (Clustered Regulatory Interspaced Short Palindromic Repeats) Cas9 system with three guide RNAs (gRNAs) was used to inactivate six homologous genes within the Bobwhite wheat genome. The resulting mutant wheat plants with all their genes disabled had slimmer stems, had a modest reduction in biomass and senesced more slowly in water limiting conditions, but did not exhibit improved yield under drought conditions. Our results show that multiplexed gRNAs enabled effective targeted gene editing of the gene family in hexaploid wheat. These mutant wheat plants will be a resource for further research studying the function of this gene family in wheat.