Scab is a devastating disease in wheat and barley. Significant 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-TSCI 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 first developed an all-in-one construct including the CRISPR/Cas9, sgRNA targeting the mlo locus, and Fhb7 donor DNA and transformed into 'Golden Promise' by Agrobacterium mediation, which produced 39 transgenic plants. Detached leaf assay showed Fhb7 function in resistance to Fusarium graminearum, but no targeted insertions were detected in the T1 populations possibly due to low copy number of the donor DNA. Subsequently, we use 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.' From about 300 T0 plants, we identified 13 Fhb7 insertion lines. The Fhb7 gene transmitted to the T1 generation. We are in the process of characterizing the insertion junctions. I will discuss the detailed progress of this TSCI project in my presentation.