Enabling High-throughput Approaches for Rapid Screening in the Age of Genome-Editing and Synthetic Biology

Significant advances in synthetic biology and genome-editing have necessitated the need for rapid screening modalities for plants, with the ultimate goal of speeding the path to the development of next-gen crops. This emerging need has brought about a resurgence in the use of protoplasts for transient screening of regulatory elements and gRNA targets. For high-throughput applications, protoplasts have several advantages over cell suspension cultures: 1) Ability to use chemical transformation systems, 2) Distinct single-cell morphology, 3) Ability to rapidly regenerate primary cell wall, and 4) Rapid expression of marker genes. Further, owing to the totipotent nature of plant cells, protoplasts “should” be capable of regeneration into whole plants. Unfortunately, where mammalian research has dedicated significant efforts into organogenesis and cell differentiation from stem cells, plant biology is still in its infancy in making similar strides with protoplasts. Leveraging recent research success, we will illustrate several examples of high-throughput applications utilizing protoplast systems in various species, and lay the foundation for advancing this important research area for the development of next-gen crops.