Photoselective sequencing: microscopically guided genomic measurements with subcellular resolution.
Photoselective sequencing: microscopically guided genomic measurements with subcellular resolution.
复制标题
光选择性测序：具有亚细胞分辨率的显微镜引导基因组测量。
DOI：
10.1038/s41592-023-01845-8
复制
发表时间：
2023
期刊：
影响因子：
48
通讯作者：
Chen,Fei
中科院分区：
文献类型：
--
作者：
Mangiameli,SarahM;Chen,Haiqi;Earl,AndrewS;Dobkin,JulieA;Lesman,Daniel;Buenrostro,JasonD;Chen,Fei
在生物系统中，空间组织和功能是相互关联的。在这里，我们提出了光选择性测序，一种新的方法，基因组和表观基因组分析在形态上不同的地区。从完整的生物样本开始，光选择性测序使用靶向照明来选择性地解除光笼化片段文库的封闭，将基于测序的读出限制在显微镜下识别的空间区域。我们通过测量小鼠脑内荧光标记细胞类型的染色质可及性概况并与已发表的数据进行比较来验证光选择性测序。此外，通过将光选择性测序与用于分解批量可访问性配置文件的计算策略相结合，我们发现少突胶质细胞谱系细胞群体相对富集皮质中的少突胶质细胞祖细胞与胼胝体。最后，我们利用光选择性测序在亚细胞规模，以确定与定位在核周边的染色质的功能。这些结果共同表明，光选择性测序是一个灵活的和可推广的平台，用于探索空间结构与基因组和表观基因组特性的相互作用。
In biological systems, spatial organization and function are interconnected. Here we present photoselective sequencing, a new method for genomic and epigenomic profiling within morphologically distinct regions. Starting with an intact biological specimen, photoselective sequencing uses targeted illumination to selectively unblock a photocaged fragment library, restricting the sequencing-based readout to microscopically identified spatial regions. We validate photoselective sequencing by measuring the chromatin accessibility profiles of fluorescently labeled cell types within the mouse brain and comparing with published data. Furthermore, by combining photoselective sequencing with a computational strategy for decomposing bulk accessibility profiles, we find that the oligodendrocyte-lineage-cell population is relatively enriched for oligodendrocyte-progenitor cells in the cortex versus the corpus callosum. Finally, we leverage photoselective sequencing at the subcellular scale to identify features of chromatin that are correlated with positioning at the nuclear periphery. These results collectively demonstrate that photoselective sequencing is a flexible and generalizable platform for exploring the interplay of spatial structures with genomic and epigenomic properties.