Department of Biological Sciences, NMIMS Sunandan Divatia School of Science, SVKM's NMIMS (Deemed to-Be University), Mumbai, Maharashtra, India.
Symbiosis Centre for Stem Cell Research (SCSCR), Symbiosis International University (SIU), Pune, Maharashtra, India.
Methods Mol Biol. 2022;2520:117-133. doi: 10.1007/7651_2021_400.
Neuronal differentiation is an intricate and a complex process which involves crosstalk among various signaling pathways, growth factors, transcription factors, and epigenetic modifiers. During different stages of neuronal development, there are various histone modifiers which drive the expression of lineage-specific genes. Polycomb group proteins are one of the histone modifiers that control transcriptional repression of specific genes in development, differentiation, and functionality of various tissues. Chromatin immunoprecipitation (ChIP) is a widely used technique to investigate the interaction of proteins and DNA; ChIP combined with quantitative real-time PCR (qPCR) gives a quantitative data about the occupancy of specific protein on a particular stretch of DNA, and this can help us investigate how a protein regulates expression of a specific gene. In this chapter, we describe a protocol for ChIP coupled to qPCR during early neuronal differentiation to identify the specific genomic targets regulated by components of Polycomb repressive complex 1.
神经元分化是一个复杂的过程，涉及各种信号通路、生长因子、转录因子和表观遗传修饰物之间的串扰。在神经元发育的不同阶段，有各种组蛋白修饰物驱动谱系特异性基因的表达。Polycomb 组蛋白是控制特定基因在发育、分化和各种组织功能中转录抑制的组蛋白修饰物之一。染色质免疫沉淀（ChIP）是一种广泛用于研究蛋白质和 DNA 相互作用的技术；ChIP 与定量实时 PCR（qPCR）结合使用，可以提供特定蛋白质在特定 DNA 片段上的占有率的定量数据，这可以帮助我们研究蛋白质如何调节特定基因的表达。在本章中，我们描述了一种在早期神经元分化过程中结合 qPCR 的 ChIP 方案，以鉴定由 Polycomb 抑制复合物 1 组成部分调控的特定基因组靶标。
