STING在神经干细胞特异敲除小鼠大脑皮层转录组及单细胞数据

为了研究STING缺失表型相关的分子机制，我们通过RNA测序(RNA-seq)分析了WT与STINGcKOE13大脑皮层的整体转录组差异。采用安捷伦2100生物分析仪(Annoroad Genomics)对总RNA进行定量和质量控制。cDNA文库构建完成后，利用Illumina HiSeq 2500平台进行高通量测序。
为了进一步明确STINGcKO大脑皮层的异常，我们使用10x Genomics平台对P0时期的大脑皮层组织进行了单细胞RNA测序(scRNA-seq)。在WT (STINGfl/fl)和STINGcKO样本：在冰上将P0的小鼠大脑皮层剥离出来，用人工脑脊液配置的Papain(20 U mg−1; Worthington)消化成单细胞悬液，37 °C ，20 min。然后细胞悬液过40 μm的滤筛，1200 rpm 5 min，弃上清。细胞用冰浴过的人工脑脊液洗细胞两次，1200 rpm 5 min。细胞用冰浴的含有1%BSA的DPBS吹悬，通过计数将细胞浓度稀释到10万细胞每毫升。最后利用安诺优达公司的10X Genomics 试剂和仪器进行测序分析。

To investigate the molecular mechanisms underlying the STING-deficient phenotype, we analyzed the global transcriptomic differences between wild-type (WT) and STINGcKOE13 cerebral cortices via RNA sequencing (RNA-seq). The total RNA was quantified and quality-controlled using an Agilent 2100 Bioanalyzer (Annoroad Genomics). After cDNA library construction, high-throughput sequencing was performed on the Illumina HiSeq 2500 platform.

To further characterize the abnormalities in STINGcKO cerebral cortices, we conducted single-cell RNA sequencing (scRNA-seq) on postnatal day 0 (P0) cerebral cortical tissues using the 10x Genomics platform. For WT (STINGfl/fl) and STINGcKO samples: P0 mouse cerebral cortices were dissected on ice, then digested into single-cell suspensions with Papain (20 U mg−1; Worthington) reconstituted in artificial cerebrospinal fluid (aCSF) at 37 °C for 20 min. The cell suspension was then passed through a 40 μm cell strainer, centrifuged at 1200 rpm for 5 min, and the supernatant was discarded. The cells were washed twice with ice-cold aCSF, centrifuged at 1200 rpm for 5 min each time. The cells were resuspended in ice-cold DPBS supplemented with 1% BSA, and the cell concentration was adjusted to 1 × 10^5 cells/mL via cell counting. Finally, sequencing analysis was performed using 10x Genomics reagents and instruments from Annoroad Genomics.

搜集汇总

背景与挑战

背景概述

该数据集包含STING条件敲除小鼠大脑皮层的转录组和单细胞RNA-seq数据，涵盖E13和P0两个发育阶段，旨在研究STING如何调控神经前体细胞的增殖和皮层发育。数据包括野生型和敲除型样本，使用Illumina平台测序，并提供了详细的分析比较文件。