Exploring the role and implications of a Histone Deacetylase III (HDAC3) and NCOR/SMRT in cancer and embryonic stem cell differentiation

探索组蛋白脱乙酰酶 III (HDAC3) 和 NCOR/SMRT 在癌症和胚胎干细胞分化中的作用和影响

• 批准号: 2264698
• 金额: --
• 依托单位: University of Leicester
• 依托单位国家: 英国
• 项目类别: Studentship
• 财政年份: 2019
• 资助国家: 英国
• 起止时间: 2019 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=studentship-2264698
• 关键词: Exploring; role; implications; Histone; Deacetylase

Histone deacetylase (HDAC) enzymes regulate global chromatin accessibility by removing the acetyl moiety from Lys residues in histone tails, restoring their positive charge and tightening chromatin to prevent gene transcription. The highly related class-I HDACs and their associated co-repressor complexes (Sin3, NuRD, CoREST, MiDAC and NCoR) have roles in many biological processes including, but not limited to, embryonic development, cell cycle regulation and apoptosis. The primary aim of this research is to create a HDAC3 with a degradation TAG (dTAG) to elucidate its role in embryonic development and cancer progression. The system uses CRISPR/Cas9 technology to incorporate a mutant FKBP12F36V tag into the gene of the protein of interest (POI), such that the resulting fusion protein (HDAC3-dTAG) can be targeted with a PROTAC molecule. Protein targeting chimeras (PROTACs) are hetero-bifunctional molecules that incorporate highly-selective E3 ligase and FKBP12F36V ligands to exploit a cells degradation system to remove the POI. The rapid degradation of HDAC3 (~2 hours) will allow us to examine its role within the same time-frame as gene regulation and DNA synthesis, something not previously possible with knockout models. We expect to extend our understanding of class-I HDAC function in cell proliferation and homeostasis that will inform their use as drug targets in numerous disease types.

组蛋白脱乙酰基酶（HDAC）通过从组蛋白尾部中的Lys残基去除乙酰基部分、恢复其正电荷并收紧染色质以阻止基因转录来调节全局染色质可及性。高度相关的I类HDAC及其相关的共阻遏物复合物（Sin 3、NuRD、CoREST、MiDAC和NCoR）在许多生物过程中具有作用，包括但不限于胚胎发育、细胞周期调节和细胞凋亡。本研究的主要目的是创建具有降解TAG（dTAG）的HDAC 3，以阐明其在胚胎发育和癌症进展中的作用。该系统使用CRISPR/Cas9技术将突变体FKBP 12 F36 V标签整合到目标蛋白（POI）的基因中，从而可以用PROTAC分子靶向所得融合蛋白（HDAC 3-dTAG）。蛋白质靶向嵌合体（PROTAC）是异源双功能分子，其并入高选择性E3连接酶和FKBP 12 F36 V配体以利用细胞降解系统来去除POI。HDAC 3的快速降解（约2小时）将使我们能够在与基因调控和DNA合成相同的时间范围内研究其作用，这在以前的敲除模型中是不可能的。我们期望扩展我们对I类HDAC在细胞增殖和稳态中的功能的理解，这将告知它们在许多疾病类型中作为药物靶点的用途。