RNA-Seq Analysis of Mutant Prss56-/- And Control Prss56+/- Retinal Transcriptomes

Method: To study the molecular processes underlying PRSS56-mediated ocular size regulation, we performed a high throughput gene expression analysis on the retina from Prss56-/- mutant mice and their control Prss56+/- littermates. Prss56+/- (Heterozygous) and Prss56+/+ (Wild type) are phenotypically similar and therefore we used heterozygous mice as controls in our experiments. Briefly, retinas from post-natal day 15 (P15) mice were extracted after enucleation, flash frozen on dry ice and RNA was isolated using Qiagen RNeasy Mini Kit as per manufacturers protocol. The RNA-Seq was performed by Novogene Corporation Inc. (Sacramento, USA) using Illumina NovaSeq 6000 Platform. Results: Our transcriptome analysis (RNA-seq) identified Adamts19 and Prss56 as the top two differentially expressed genes between Prss56 mutant (Prss56-/-) and control (Prss56+/-) in P15 retina. Retinal mRNA profiles of 15-day old wild type (Prss56+/-) and Prss56-/- mice.研究方法：为解析PRSS56介导的眼球大小调控的分子过程，我们对Prss56基因敲除（Prss56-/-）突变小鼠及其同窝杂合对照（Prss56+/-）的视网膜开展了高通量基因表达分析。Prss56+/-（杂合子）与Prss56+/+（野生型）表型高度相似，故本实验以杂合小鼠作为对照。简言之，我们于小鼠出生后第15天（P15）摘除眼球后分离视网膜，置于干冰中快速冷冻，并依照制造商说明书使用Qiagen RNeasy Mini试剂盒提取总RNA。RNA测序（RNA-Seq）由美国萨克拉门托的诺沃基因公司（Novogene Corporation Inc.）依托Illumina NovaSeq 6000测序平台完成。 研究结果：本转录组分析（RNA-seq）鉴定出，在P15天小鼠视网膜中，Adamts19与Prss56为Prss56突变型（Prss56-/-）与对照（Prss56+/-）组间差异表达最显著的两个基因。本数据集包含15日龄野生型（Prss56+/-）与Prss56-/-小鼠的视网膜mRNA表达谱。