Functional genetic screens for structural regulatory DNA elements driving tumor progression and drug resistance

驱动肿瘤进展和耐药性的结构调节 DNA 元件的功能遗传筛选

• 批准号: 415497046
• 负责人: Dr. Sebastian Dieter
• 金额: --
• 依托单位: Nationales Centrum für Tumorerkrankungen (NCT) Heidelberg – DKFZ
• 依托单位国家: 德国
• 项目类别: Research Fellowships
• 财政年份: 2018
• 资助国家: 德国
• 起止时间: 2017-12-31 至 2020-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/415497046?language=en
• 关键词: Functional; genetic; screens; structural; regulatory

The human genome carries genetic information in two distinct forms: Transcribed coding and non-coding genes as well as non-transcribed regulatory DNA elements (rDEs). rDEs control the magnitude and pattern of expression of genes and are indispensable for organismal development. In line with this, increasing evidence indicates that variation of rDEs is key to tumor pathogenesis, suggesting that rDEs play crucial roles in cell identity in health and disease.rDEs can be subdivided into transcriptional rDEs (t-rDEs) that recruit transcription factors to enhancers, thereby affecting gene expression, and structural rDEs (s-rDEs) that maintain chromatin 3D structure, a feature essential for regulation of genes by t-rDEs. While large-scale functional genetic screens greatly advanced our knowledge in studying mammalian genes, such tools to analyze rDEs were lacking. In the last few years, spectacular advance in genome editing technologies by the CRISPR/Cas system has not only expanded the available screening toolbox to examine genes, but also opened up novel possibilities for studying rDEs. Recently, the Agami group has developed a CRISPR strategy to target t-rDEs and has shown its scalability and effectivity in identifying potential oncogenic and tumor-suppressive t-rDEs. In the proposed project, we will exploit this experience for targeting s-rDEs, and setup functional genetic screens to identify key players in cell proliferation and survival, cancer progression and resistance to therapy. Moreover, we will investigate underlying molecular mechanisms. Thereby, our results will provide the functional knowledge necessary to better understand the role of s-rDEs in controlling the 3D chromatin structure in cancer and its implication for cancer progression and drug resistance.

人类基因组以两种不同形式携带遗传信息：转录的编码基因和非编码基因以及非转录的调控DNA元件（rDE）。 rDE 控制基因表达的大小和模式，对于生物体发育是不可或缺的。与此相一致的是，越来越多的证据表明，rDE 的变异是肿瘤发病机制的关键，这表明 rDE 在健康和疾病中的细胞身份中发挥着至关重要的作用。rDE 可以细分为转录 rDE (t-rDE)，其招募转录因子以增强子，从而影响基因表达；以及结构 rDE (s-rDE)，其维持染色质 3D 结构，这是基因调控所必需的特征 通过 t-rDE。虽然大规模的功能遗传筛选极大地提高了我们研究哺乳动物基因的知识，但我们仍然缺乏分析 rDE 的工具。在过去几年中，CRISPR/Cas 系统在基因组编辑技术方面取得了惊人的进步，不仅扩展了用于检查基因的可用筛选工具箱，而且还为研究 rDE 开辟了新的可能性。最近，Agami 小组开发了一种针对 t-rDE 的 CRISPR 策略，并显示了其在识别潜在致癌和肿瘤抑制 t-rDE 方面的可扩展性和有效性。在拟议的项目中，我们将利用这一经验来靶向 s-rDE，并设置功能遗传筛选来识别细胞增殖和存活、癌症进展和治疗耐药性中的关键参与者。此外，我们将研究潜在的分子机制。因此，我们的结果将提供必要的功能知识，以更好地理解 s-rDE 在控制癌症 3D 染色质结构中的作用及其对癌症进展和耐药性的影响。