Primary　organoid　cultures　generated　from　patient　biopsies　comprise　a　novel　improved　platform　for　disease　modeling,　being　genetically　stable　and　closely　recapitulating　in　vivo　scenarios.　Barrett　esophagus　(BE)　is　the　major　risk　factor　for　esophageal　adenocarcinoma.　There　has　been　a　dearth　of　long-term　in　vitro　expansion　models　of　BE　neoplastic　transformation.　We　generated　a　long-term　virus-free　organoid　expansion　model　of　BE　neoplasia　from　patient　biopsies.　Both　wild-type　and　paired　APC-knockout　(APC(KO))　BE　organoids　genome-edited　by　CRISPR-Cas9　showed　characteristic　goblet　cell　differentiation.　Autonomous　Wnt　activation　was　confirmed　in　APC(KO)　organoids　by　overexpression　of　Wnt　target　genes　and　nuclear-translocated　beta-catenin　expression　after　withdrawal　of　Wnt-3A　and　R-spondin-1.　Wnt-activated　organoids　demonstrated　histologic　atypia,　higher　proliferative　and　replicative　activity,　reduced　apoptosis,　and　prolonged　culturability.　Wnt-activated　organoids　also　showed　sustained　protrusive　migration　ability　accompanied　by　disrupted　basement　membrane　reorganization　and　integrity.　This　CRISPR-Cas9　editing　human-derived　organoid　model　recapitulates　the　critical　role　of　aberrant　Wnt/beta-catenin　signaling　activation　in　BE　neoplastic　transformation.　This　system　can　be　used　to　study　other　＇driver＇　pathway　alterations　in　BE-associated　neoplasia,　avoiding　signaling　noise　present　in　immortalized　or　cancer-derived　cell　lines.