The Barley Genetic Engineering Facility accepts requests from barley researchers to develop tissue culture, transformation, and regeneration protocols for their chosen barley cultivars. Once a successful tissue culture protocol is established, the lab will accept requests for gene transformation experiments using the specific barley cultivar. Confirmed T1 transgenic barley seeds will be delivered to the requesting researchers, along with the USDA-APHIS permits that they provide.
Currently, successful transformation and regeneration protocols have been developed for the two-rowed cvs. ‘Conlon’ and ‘Genesis’ and the six-rowed cv. ‘Morex’ using immature scutellum as the explant. Transgenic barley plants can be produced via both biolistic bombardment and Agrobacterium-mediated transformation. The selectable marker is the hygromycin resistance gene. With the support of funding from the USWBSI, the laboratory of Drs. Di and Lawton has developed CRISPR-gene editing platforms for barley with single-guide RNA to knock-out a gene and dual/multiple tRNA design for multiplexed gene editing. The Barley Genetic Engineering Facility provides guidance on CRISPR-gene editing vector designing and accepts gene editing requests from barley researchers.
Contact: Dr. Rong Di | Barley Submission Form | Barley Transformation Lab Presentation
The Wheat Transformation/Gene Editing Facility accepts transformation and gene editing requests from researchers working on FHB-related research. Transformation requests will be processed in order of receipt; however, clients active with the USWBSI and those requesting agronomically adapted germplasm would have priority. Researchers without an active USWBSI grant but working on FHB-related research would need to get approval from the Gene Discovery and Engineering Resistance Research Committee. Clients would provide vectors for transformation/gene editing.
Three to five months after the initiation of a project putative events will be screened for glufosinate resistance by herbicide painting and then DNA samples will be collected for PCR analyses for the presence of the transgenes. PCR testing for the bar gene will occur at KSU, and testing for the gene of interest (GOI) will be performed either at KSU or by the individual PI as predetermined during consultation. Transgenic events will be grown to maturity at KSU and T1 generation seeds will be harvested, packed, and shipped to clients for downstream analyses. Clients would expect delivery of seeds starting approximately six months from project initiation. Prior to shipment clients would be responsible for obtaining an APHIS permit for movement of regulated articles.
Contact: Dr. Harold Trick | Wheat Submission Form | Wheat Transformation Lab Webinar
