GEOapplication/xmlftp://ftp.ncbi.nlm.nih.gov/geo/series/GSE292nnn/GSE292882/primary200OKOther Homo sapiensunidentifiedOtherhttps://www.ncbi.nlm.nih.gov/geo/query/acc.cgi?acc=GSE292882GEOGSEfalseFANCM is Required for the PAX3::FOXO1-Driven Oncogenic Program in Rhabdomyosarcoma [CRISPR Screen]Alveolar rhabdomyosarcoma (ARMS) is a highly aggressive pediatric soft tissue sarcoma driven by the PAX3::FOXO1 fusion protein. Despite multipronged treatment approaches, survival rates for patients with fusion-positive (FP) ARMS remain poor, underscoring the need for novel therapeutic strategies. Here, we employ sequential multimodal CRISPR/Cas9 genetic screens to identify essential genes in FP ARMS, revealing FANCM, a key component of the Fanconi Anemia pathway, as a critical dependency. FANCM loss selectively impairs FP ARMS cell growth in vitro and in vivo, reduces PAX3::FOXO1 levels, and induces myogenic differentiation. Mechanistically, FANCM depletion exacerbates replication stress, particularly at PAX3::FOXO1 target gene loci, which leads to ATR/ATM-mediated DNA damage signaling and selective downregulation of the fusion-driven oncogenic program. CRISPR exon-tiling screens highlighted FANCM's helicase and DNA-binding domains as essential for its FP-specific dependency, linking FANCM-mediated replication fork binding and downstream ATR signaling to FP ARMS survival. This study positions FANCM as a promising therapeutic target in FP ARMS and introduces a novel paradigm for exploiting replication stress vulnerabilities in oncofusion-driven malignancies.2026/08/04GSE292882GSM8868010GSM8868011GSM8868012GSM8868013GSM8868014GSM8868015GSM8868016GSM8868017GSM8868018GSM8868019GSM8868020GSM8868021GSM8868022GSM8868023GSM8868024GSM8868025GSM8868026GSM8868027GSM8868028GSM8868029GSM8868030GSM8867980GSM8867989GSM8867981GSM8867982GSM8867983GSM8867984GSM8867985GSM8867986GSM8867987GSM8867988GSM8868000GSM8867990GSM8868001GSM8868002GSM8867991GSM8868003GSM8867992GSM8868004GSM8867993GSM8868005GSM8867994GSM8868006GSM8867995GSM8868007GSM8867996GSM8867997GSM8868008GSM8868009GSM8867998GSM8867999308823237124676292882 Homo sapiensunidentified