Variation in shoot architecture, or tillering, is an important adaptive trait targeted during the domestication of crops. A well-known regulatory factor in shoot architecture is TEOSINTE BRANCHED 1 (TB1). TB1 and its orthologs have a conserved function in integrating environmental signals to regulate axillary branching or tillering in cereals. The barley (Hordeum vulgare) ortholog of TB1, VULGARE ROW-TYPE SIX 5 (VRS5), regulates tillering and is involved in regulating row-type by inhibiting lateral spikelet development. These discoveries predominantly come from genetic studies; however, how VRS5 regulates these processes on a molecular level remains largely unknown. By combining transcriptome analysis between the vrs5 mutant and the wild-type at different developmental stages and DAP-sequencing to locate the genome-wide DNA binding sites of VRS5, we identified bona fide targets of VRS5. We found that VRS5 targets abscisic acid-related genes, potentially to inhibit tillering in a conserved way. Later in inflorescence development, VRS5 also targets row-type gene VRS1 and several known floral development genes, such as MIKCc-type MADS-box genes. This study identifies several genes for mutational analysis, representing a selection of bona fide targets that will contribute to a deeper understanding of the VRS5 network and its role in shaping barley development.