Interleukin-17 activity contributes to severe glial pathology in HD knock-in pig model

The striatum from WT and HD KI mice and pigs, and WT monkey were isolated for RNA-Seq. Total RNA was extracted using TRIzol Reagent according to the manufacturer’s protocol. Construction of the cDNA library and sequencing were performed by Shenzen acegen Company. Clean reads were obtained after discarding reads containing adapter, poly-N or low-quality raw data. High-quality reads were aligned to the existing reference genome. Next, the HTseq was used to convert aligned short reads into read counts. DEGs were chosen according to the criteria of a fold change of greater than 1.5 and an adjusted P value of less than 0.05.

本研究分离野生型（Wild Type, WT）、亨廷顿病敲入（Huntingtin Knock-In, HD KI）小鼠、猪以及野生型猴的纹状体组织，用于RNA测序（RNA-Seq）实验。按照试剂制造商提供的操作流程，使用TRIzol试剂提取总RNA。cDNA文库构建及测序由深圳acegen公司完成。过滤掉包含接头序列、poly-N序列或低质量的原始测序读段后，得到清洁读段（clean reads）。将高质量读段比对至现有参考基因组。随后，使用HTseq工具将比对后的短读段转换为读段计数（read counts）。差异表达基因（Differentially Expressed Genes, DEGs）的筛选标准为：折叠变化大于1.5，且校正后P值小于0.05。