重叠的结合点是TF基因组占用的基础
bioRxiv : the preprint server for biology
|March 18, 2024
概括
我们开发了PADIT-seq来检测低亲和力转录因子 (TF) -DNA相互作用,揭示了重叠的结合位如何影响体内基因调节. 这种方法扩展了对影响TF结合的非编码变体的搜索.
科学领域:
- 分子生物学分子生物学
- 基因组学就是基因组学.
- 表观遗传学 在表观遗传学中,表观遗传学是指表观遗传学.
背景情况:
- 转录因子 (TFs) 通过特定序列的DNA结合来调节基因表达.
- 目前的高通量方法难以检测基因调节至关重要的低亲和度TF-DNA相互作用.
研究的目的:
- 开发一种敏感的方法来测试所有可能的DNA序列中的TF结合偏好.
- 描述低亲和度TF-DNA相互作用及其在TF基因组占用中的作用.
主要方法:
- 开发了PADIT-seq (通过体外转录和RNA测序对DNA的蛋白亲和力) 用于高灵敏度TF结合试验.
- 对人类TFs HOXD13和EGR1.1的所有10bpDNA序列进行测试TF结合.
主要成果:
- PADIT-seq检测到TF绑定偏好比以前的方法更灵敏.
- 确定了HOXD13和EGR1.1的低亲和度DNA结合点的广泛目录.
- 揭示了侧边核酸形成重叠的,低亲和度的网站,调节TF结合和基因组占用 in vivo.
结论:
- PADIT-seq为检测TF-DNA相互作用提供了前所未有的灵敏度.
- 了解重叠的结合点是解读TF基因组占用的关键.
- 这项工作扩大了基因组序列空间,用于识别影响TF结合和基因调节的非编码变异.
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