Department of Life Sciences, Korea University, Seoul, South Korea.
Methods Mol Biol. 2021;2261:345-356. doi: 10.1007/978-1-0716-1186-9_21.
In plant cells, transcription factors play an important role in the regulation of gene expression, which eventually leads to the formation of complex phenotypes. Although chromatin immunoprecipitation (ChIP) involves a lengthy process that requires up to 4 days to complete, it is a powerful technique to investigate the interactions between transcription factors and their target sequences in vivo. Here, we describe a detailed ChIP protocol, focusing on ChIP-qPCR, from material collection to data analyses. Moreover, we explain multiple checkpoints for the quality control of ChIP-qPCR data to ensure the success of this protocol. As this protocol is robust, it can be adapted to other plant materials and plant species, and it can be used for genome-wide profiling experiments, including ChIP-chip and ChIP-seq analyses. We believe that our ChIP-qPCR protocol facilitates research on the interactions between plant transcription factors and their target sequences in vivo.
在植物细胞中,转录因子在基因表达的调控中起着重要作用,最终导致复杂表型的形成。尽管染色质免疫沉淀(ChIP)涉及一个需要长达 4 天才能完成的冗长过程,但它是一种强大的技术,可以研究转录因子与其靶序列在体内的相互作用。在这里,我们描述了一个详细的 ChIP 方案,重点是 ChIP-qPCR,从材料收集到数据分析。此外,我们还解释了 ChIP-qPCR 数据质量控制的多个检查点,以确保该方案的成功。由于该方案具有稳健性,因此可以将其应用于其他植物材料和植物物种,并且可以用于全基因组分析实验,包括 ChIP-chip 和 ChIP-seq 分析。我们相信,我们的 ChIP-qPCR 方案有助于研究植物转录因子与其靶序列在体内的相互作用。