The intestinal immune system must maintain tolerance to harmless dietary and microbial antigens while preserving protective immunity against pathogens. Peripherally induced regulatory T (pTreg) cells play a pivotal role in mediating this tolerance. We recently identified PRDM16⁺ tolerogenic dendritic cells (tolDCs) as a distinct antigen-presenting cell population required for the differentiation of both food- and microbiota-specific pTreg cells. Using multiple genetic models, we found that disruption of the tolerogenic DCs through deletion of Prdm16, Rorc(t), or the Rorc(t)+7 kb cis-regulatory element resulted in microbiota-specific proinflammatory T cell responses and a breakdown of oral tolerance, rendering animals susceptible to allergic inflammation. We next investigated the developmental origin and persistence of this population. PRDM16+ tolDCs were labeled by both Il7r- and Csf1r-based fate-mapping approaches, and disruption of either pathway alone was insufficient to eliminate their development or function. Notably, PRDM16+ tolDC development and function were independent of common γ chain signaling. Furthermore, inducible depletion of PRDM16+ tolDCs in adult mice abolished the generation of food antigen-specific pTreg cells. The PRDM16+ tolDC population subsequently recovered, together with its capacity to induce pTreg cell differentiation. To selectively label PRDM16+ tolDCs in vivo, we generated a Prdm16/Rorc(t) intersectional reporter mouse, providing a new genetic tool for investigating this specialized DC population. Together, these findings establish PRDM16+ tolDCs as a central regulator of intestinal immune tolerance and provide new insights into their development and maintenance.