胶质母细胞瘤（GBM）干细胞样细胞（GSCs）对于GBM的发生和治疗抗药性至关重要。越来越多的证据支持长链非编码RNA（lncRNAs）具有促进肿瘤功能，但它们在GSCs中的作用仍不明确。通过将CRISPRi筛选与多组学方法结合，我们发现了一个由lncRNA DARS1-AS1控制的转录后调控回路，促进了GBM细胞/GSCs的恶性特性。DARS1-AS1的耗竭抑制了GBM细胞/GSCs的增殖和GSCs的自我更新，延长了原位GBM模型的生存时间。DARS1-AS1耗竭还削弱了同源重组（HR）介导的双链断裂（DSB）修复，并增强了GBM细胞/GSCs的放射敏感性。在机制上，DARS1-AS1与YBX1相互作用，促进目标mRNA的结合和稳定化，形成一个混合的转录后调控正反馈环，上调了G1-S转换的关键调控因子的表达，包括E2F1和CCND1。DARS1-AS1/YBX1还稳定了FOXM1的mRNA，这是一个调控GSC自我更新和DSB修复的主要转录因子。我们的发现表明，DARS1-AS1/YBX1轴可能是GBM对放射治疗/HR缺陷靶向治疗敏感化的潜在治疗靶点。

The glioblastoma (GBM) stem cell-like cells (GSCs) are critical for tumorigenesis/therapeutic resistance of GBM. Mounting evidence supports tumor-promoting function of long noncoding RNAs (lncRNAs), but their role in GSCs remains poorly understood. By combining CRISPRi screen with orthogonal multiomics approaches, we identified a lncRNA DARS1-AS1-controlled posttranscriptional circuitry that promoted the malignant properties of GBM cells/GSCs. Depleting DARS1-AS1 inhibited the proliferation of GBM cells/GSCs and self-renewal of GSCs, prolonging survival in orthotopic GBM models. DARS1-AS1 depletion also impaired the homologous recombination (HR)-mediated double-strand break (DSB) repair and enhanced the radiosensitivity of GBM cells/GSCs. Mechanistically, DARS1-AS1 interacted with YBX1 to promote target mRNA binding and stabilization, forming a mixed transcriptional/posttranscriptional feed-forward loop to up-regulate expression of the key regulators of G1-S transition, including E2F1 and CCND1. DARS1-AS1/YBX1 also stabilized the mRNA of FOXM1, a master transcription factor regulating GSC self-renewal and DSB repair. Our findings suggest DARS1-AS1/YBX1 axis as a potential therapeutic target for sensitizing GBM to radiation/HR deficiency-targeted therapy.