In vivo tissue-specific chromatin profiling in Drosophila melanogaster using GFP-tagged nuclei
In vivo tissue-specific chromatin profiling in Drosophila melanogaster using GFP-tagged nuclei
复制标题
DOI：
10.1101/2021.03.23.436625
复制
发表时间：
2021-03
期刊：
影响因子：
--
通讯作者：
Juan Jauregui-Lozano;Kimaya M. Bakhle;Vikki M. Weake
中科院分区：
文献类型：
--
作者：
Juan Jauregui-Lozano;Kimaya M. Bakhle;Vikki M. Weake
染色质景观定义了多细胞生物体中的细胞身份，具有独特的DNA可及性模式和装饰每种细胞类型基因组的组蛋白标记。因此，分析疾病或生理条件下完整生物体中不同细胞类型的染色质状态可以提供对染色质如何调节体内细胞稳态的洞察。为了克服与表征特定细胞类型中的染色质状态相关的许多挑战，我们开发了一种改进的方法来分离在Gal4/UAS控制下表达的GFP标记的果蝇核。使用该方案，我们使用Omni-ATAC分析了染色质可及性，并使用ChIP-seq和CUT & Tag检查了成年感光神经元中组蛋白标记的分布。我们发现，光感受器的染色质景观反映了这些细胞的转录状态，证明了我们的方法的质量和再现性，用于分析果蝇中特定细胞类型的转录组和表观基因组。
The chromatin landscape defines cellular identity in multicellular organisms with unique patterns of DNA accessibility and histone marks decorating the genome of each cell type. Thus, profiling the chromatin state of different cell types in an intact organism under disease or physiological conditions can provide insight into how chromatin regulates cell homeostasis in vivo. To overcome the many challenges associated with characterizing chromatin state in specific cell types, we developed an improved approach to isolate Drosophila nuclei tagged with GFP expressed under Gal4/UAS control. Using this protocol, we profiled chromatin accessibility using Omni-ATAC, and examined the distribution of histone marks using ChIP-seq and CUT&Tag in adult photoreceptor neurons. We show that the chromatin landscape of photoreceptors reflects the transcriptional state of these cells, demonstrating the quality and reproducibility of our approach for profiling the transcriptome and epigenome of specific cell types in Drosophila.