In Situ Hybridization for Detecting Mature MicroRNAs In Vivo at Single-Cell Resolution.
In Situ Hybridization for Detecting Mature MicroRNAs In Vivo at Single-Cell Resolution.
复制标题
用于以单细胞分辨率检测体内成熟 MicroRNA 的原位杂交。
DOI：
10.1002/cpmb.93
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发表时间：
2019
影响因子：
--
通讯作者：
Arur,Swathi
中科院分区：
文献类型：
--
作者：
Minogue,AmandaL;Arur,Swathi
MicroRNAs (miRNAs) are key regulators of cell and tissue development. However, spatial resolution of miRNA heterogeneity and accumulation patternsin vivoremains uncharted. Next‐generation sequencing methods assay miRNA abundance in tissues, yet these analyses do not provide spatial resolution. A method to assay miRNA expression at single‐cell resolutionin vivoshould clarify the cell‐autonomous functions of miRNAs, their roles in influencing the cellular microenvironment, and their perdurance and turnover rate. We present anin situhybridization protocol to map miRNA subcellular expression in single cellsin vivoin four days. Using this protocol, we mapped distinct miRNAs that accumulate in the cytoplasm of one sibling oocyte but not another, dependent on the oocyte developmental stage. Thus, this method provides spatial and temporal resolution of the heterogeneity in expression of miRNAs duringCaenorhabditis elegansoogenesis. This protocol can generally be adapted to any tissue amenable to dissection and fixation. © 2019 by John Wiley & Sons, Inc.
