Opportunities and Obstacles for Candidate Gene Validation Using Crispr/Cas in Soybean.

There is considerable interest in using gene editing technologies for gene function studies and genetic improvement of crop species. This talk will address our recent findings in soybean mutagenesis using a CRISPR/Cas9-based approach.  To date, we have used CRISPR/Cas9 to induce targeted mutations in whole soybean plants for six different endogenous genes, including both in-frame and frameshift alleles.  T0 plants exhibited a high rate of mutagenesis, including multi-allelic tissue samples, chimerism within individual plants, and simultaneous mutagenesis of paralogous genes.  However, inheritance patterns of both the CRISPR/Cas9 transgenes and targeted mutations were inconsistent among the different events.  These results provide insight into the opportunities and potential obstacles that exist in identifying and stabilizing heritable soybean mutations using our CRISPR/Cas9 approach. The findings also offer important considerations for how gene editing can be used to assess candidate genes that underlie quantitative trait loci, and how these techniques may be used to introgress alleles into breeding materials in the future.