Soybean is a desired global commodity due its high-value oil and protein reserves, which make up approximately 20 and 40 percent of the seed dry weight, respectively. The ratio of these reserves is mainly controlled by genetics. Modern soybean breeding programs have successfully selected for yield, but a byproduct of this effort has been a reduction in protein and an increase in oil in the harvested seed. This project is designed to lay the foundation needed to break the linkage between protein/oil/yield. Protein re-balancing occurs during seed development when common seed-specific proteins are disrupted or lost. The outcome is the increased expression of other genes to compensate for the depletion in the major proteins that the seed will require for germination, a common phenomenon in plant species. Depending on which proteins compensate for the loss of the major protein reserve, the final protein composition can influence nutritional and/or functionality qualities of the resulting seed protein content, without compromising seed oil or yield. Our team has employed genome editing approaches to create null mutations in major soybean seed protein genes, including members of the glycinin and beta-conglycinin gene families, which triggered protein re-balancing. In addition, a set of edits were created in the gene that underlies the major protein QTL on chromosome 20, cqSeed protein-003, and its paralog on chromosome 10. These genetic resources were developed, in part, from previous rounds of USB/QSSB funding. The various gene edits reside in five different soybean varieties. These rebalanced plants, and edited cqSeed protein-003 allele and paralog, were investigated, under diverse field environments. The goal was to assess changes in seed reserve content in edited lines across multiple environments and monitor impacts on the nutritional value and functionality of the seed reserves, and to use this information to gain insight on the genetic underpinnings that govern final seed reserve (oil/protein/starch) content in soybean.