Genetic engineering of plants is critical for crop improvement and requires regeneration of whole plants from genetically modified cells. While all plant species possess genetic components to form whole plants from single cells, many crop plants, including maize, have a limited ability to regenerate from cells that are subjected to genetic modification. Master genes are hypothesized to be involved in the process of plant regeneration. This is strongly related to the ability of cell proliferation, an increase in the number of cells as a result of growing and multiplying of cells. In this project, maize genes responsible for controlling cell growth and multiplication will be identified using a novel high-throughput screening approach. Validating key candidate genes in regulating maize regeneration will be carried out through further testing. The primary goal of this project is to better understand the genetic basis of plant regeneration for improved genetic engineering strategies. This project will provide new ways of analyzing and understanding plant genetics through the use of new genome editing technologies, large amounts of data, and computational tools. It will also offer training opportunities in various fields such as plant genetics, genomics, molecular biology, genome editing, bioinformatics, and machine learning.<br/><br/>Plant transformation and regeneration are key procedures of genome engineering. The overarching goal of this proposal is to improve our understanding of the genetic basis of plant regeneration. As each plant genome possesses totipotent components to form an entire plant but the capability to regenerate is at low levels in many crop species, it is hypothesized that conserved master genes for totipotency governing embryogenesis exist and that additional non-conserved genes account for differing regeneration ability, and these genes are associated with the ability of cell proliferation and growth. In this project, maize genes controlling cell proliferation and growth will be identified by conducting analysis of transcriptomic gene networks, genomic comparisons, high-throughput CRISPR screening, and functional validation of individual genes. The project will provide innovative approaches, massive data, and computational tools for integrative genomic analysis and discoveries. T<br/><br/>This project is jointly funded by the Plant Genome Research Program in the Directorate for Biological Sciences and the Established Program to Stimulate Competitive Research (EPSCoR).<br/><br/>This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.