Scab is a devastating disease in wheat barley. Considerable progress has been made in understanding and improving host resistance in wheat with molecular cloning of the major QTL Fhb1 and Fhb7; however, similar research with barley has lagged due to the lack of highly resistant genotypes, which makes it difficult to effectively control FHB and DON contamination. Supported by the USWBSI-TRSC program, we are developing marker-free transfer of Fhb7, encoding a glutathione S-transferase functioning in the detoxification of mycotoxins including DON, to barley via CRISPR-mediated targeted gene insertion. We used chemically modified Fhb7 donor DNA and a CRISPR–Cas9 construct targeting the mlo locus to transform the immature embryos of the elite two-rowed malting barley cultivar Excelsior Gold (EG). From about 300 T₀ plants, we identified 13 Fhb7 insertion lines. We also developed 20 transgenic plants in Gold Promise (GP) carrying two T-DNA cassettes, one for Fhb7 gene and another for transformation selection marker HygR by Agrobacterium mediation. We have obtained marker-free Fhb7 transgenic plants in EG and GP in the T₁ generation. Detached leaf assay showed Fhb7 function in resistance to Fusarium graminearum. Two EG transgenic lines together with non-transgenic EG as a control were tested for FHB resistance in the field and greenhouse conditions. The results showed that Fhb7 reduced FHB serenity by up to 64%. We are in the process of characterizing the insertion junctions and engineering Fhb7 for new alleles.