RNAseq analysis of retinal microglia during early diabetes

We report the use of RNAseq to identify gene changes in isolated retinal microglia after 4 weeks of STZ-induced diabetes. At this early time point in diabetes induction, single retinal cell suspensions were sorted via FACS using a Cd11b-FITC conjugate. RNA was isolated and preamplified using the SMARTSeq v4 low RNA input kit. After library preparation a 50bp single end read was performed at a depth of 19-34 million reads per sample using the Illumina HiSeq. Constructs were mapped to the rat genome and differential expression calculated between the STZ-treated and control groups. Examination of 2 groups (control and diabetic) after 4 weeks of STZ induced diabetes

本研究报道了利用RNA测序（RNAseq）鉴定链脲佐菌素（STZ）诱导糖尿病4周后分离的视网膜小胶质细胞的基因表达变化。在糖尿病诱导的该早期时间节点，通过CD11b-异硫氰酸荧光素（FITC）结合物完成视网膜单细胞悬液的荧光激活细胞分选术（FACS）分选。提取RNA后，采用SMARTSeq v4低起始量RNA试剂盒进行预扩增。完成文库构建后，使用Illumina HiSeq测序平台对每个样本开展1900万至3400万条读长的50bp单端测序。将测序读段比对至大鼠基因组，并计算STZ处理组与对照组的差异表达基因。本研究的分析对象为STZ诱导糖尿病4周后的两组样本：对照组与糖尿病组。