To explore the influence of genetics on homeostatic regulation of dendritic cell (DC) numbers, we present a screen of DCs and their progenitors in lymphoid and non-lymphoid tissues in Collaborative Cross (CC) and Diversity Outbred (DO) mice. We report 30 and 71 loci with logarithm of the odds (LOD) scores >8.18 and ranging from 6.67 to 8.19, respectively. The analysis reveals the highly polygenic and pleiotropic architecture of this complex trait, including many of the previously identified genetic regulators of DC development and maturation. Two SNPs in genes potentially underlying variation in DC homeostasis, a splice variant in Gramd4 (rs235532740) and a missense variant in Orai3 (rs216659754), are confirmed by gene editing using CRISPR-Cas9. Gramd4 is a central regulator of DC homeostasis that impacts the entire DC lineage, and Orai3 regulates cDC2 numbers in tissues. Overall, the data reveal a large number of candidate genes regulating DC homeostasis in vivo. Display omitted •Collaborative Cross and Diversity Outbred mice show dendritic cell (DC) frequency variation•QTL mapping reveals the highly polygenic and pleiotropic architecture of DC homeostasis•Gramd4 and Orai3 regulate DC frequency Oliveira et al. use quantitative trait locus (QTL) mapping in Diversity Outbred mice to find candidate genes linked to dendritic cell (DC) homeostasis. Site-directed mutagenesis using CRISPR-Cas9 confirms two candidate genes, Gramd4 and Orai3. Overall, the data represent a resource for interrogating the mechanisms governing DC homeostasis in tissues.