Gene editing (GE) has emerged as a disruptive technology that can be used to generate novel variation in the genomic regions affecting major agronomic traits. Genetic studies in cereal crops, fueled by the availability of sequenced genomes, and by the advances in sequencing technologies, identified a number of genes that control major agronomic traits in many crops. The recent release of the wheat genome sequence now provides the opportunity to easily extrapolate gene mapping information from other crops to wheat. Our project explores the capabilities of the gene editing technology to unlock the yield potential of the complex wheat genome, and build a foundation for transformative approaches to wheat improvement. We used the wheat orthologs of genes that were shown to affect yield component traits in various crops to select targets for gene editing. The gene editing pipeline based on the Cas9 and Cpf1 nucleases, and including procedures for the high-throughput screening of the designed gRNAs for editing efficiency using the wheat protoplast assays, quick assessment of the frequency and types of editing events by next-generation sequencing, and multiplex gene editing construct assembly based on the Golden Gate strategy was established for wheat. For eighteen genes, the Cas9 and Cpf1 constructs were designed and successfully tested for editing efficiency using the protoplast assay and nextgeneration sequencing. GE plants were obtained for majority of these genes in spring and winter wheat. The phenotypic effects of GE on yield component traits were confirmed for several genes, and phenotyping of remaining GE plants is underway. Wheat lines combining several edited genes were created by crossing single-gene mutants or by the modification of multiple genes and their homoeologs using a multiplex GE construct. The latter was shown can be facilitated by CRISPR/Cas9 activity maintained across generations. For example, GE showed that TaGW2 gene homoeologs’ effects are dosage-dependent and cultivar-specific with 16-20% increase in grain weight and size in triplegenome mutants. The GE variants of the seed size increasing gene transferred to other wheat cultivars showed similar phenotypic effects. An approach for high-efficiency gene editing and inter-cultivar trait transfer was established using a wheat line expressing Cas9 at the high level. In our project, we successfully demonstrated that GE technology can be used for creating novel variation in the genes that directly control yield component traits thereby building new genomic and biotechnological tools and resources for accelerating wheat improvement.