The Kausch Lab is well known in the field of plant transformation and genome editing. The lab has established a reliable pipeline of stalwart protocols for the Agrobacterium-mediated introduction of experimental genetic constructs into Sorghum cv BTx430 and collaborated to generate the viable transgenics required for the ongoing investigations on this project. The Kausch Lab functions as a central hub working with each lab, and each project accordingly, to create the stable lines necessary for this project and has many transgenic lines and events in various stages of regeneration for our initial constructs. Experiments in process through the various labs in this project include: (1) sorghum RNAi constructs for knockdowns such as for voltage-gated chloride channel proteins, alpha carbonic anhydrase 7 (CA) and nine-cis-epoxycarotenoid dioxygenase 4, and myb domain protein 60; (2) constructs to test the fidelity of phosphoenol pyruvate carboxylase (PEPC) promoter expression, CA overexpression and PEPC with altered kinetics; (3) additional versions of CA overexpression aimed to test a range of increased mesophyll CA activity; (4) Ta Cas 9, dTa Cas9, and, dCas9 transcriptional activator for improved editing, and; (5) constructs to evaluate improvements to the transgenic process with the intent to increase transformation frequencies and shorten the time to T1 seed.
