Profiling of miRNAs expression in Rag2 KO and wild type mice spleen by miRNA expression microarray analysis. Mus musculus

Purpose: miRNAs are important fators that are involved in the regulation of at least one third of the total human genes and are involved in the regulation of different biological processes. The purpose of the study was to investigate the differential expression pattern of miRNAs in Rag2 KO mice spleen compared to its wild type counterpart. Methods: In this study, we have excised the spleen tissues from three Rag2 KO mice and three wild type mice, and then RNA samples were prepared with the spleen tissues for the analysis of miRNAs profiles using the Affymetrix Genechip miRNA 4.0 arrays. The Affymetrix Genechip miRNA 4.0 array offers an updated content without compromising the high performance as the previous-generation arrays, it also provides a comprehensive coverage that is designed to interrogate all mature miRNA sequences in miRBase Release 20, as well as an easily correlate miRNA results having analysis files contain host gene ID, predicted and validated miRNA target genes, and clustered miRNA information. Result: The miRNAs expression microarray analysis showed that many miRNAs have been expressed differentially between Rag2 KO and wild type mice spleen. These miRNAs might be involved in the different biological and physiological processes including immune regulations. Conclusion: miRNAs might be an active player during theprocess of immune regulation. Overall design: Spleen miRNAs derived from Rag2 KO and wild type mice were profiled by microarray, in triplicates for each sample, by Affymetrix Genechip miRNA 4.0 array platforms.

微小RNA（miRNAs）是一类关键调控因子，可调控至少三分之一的人类全基因表达，并参与多种生物学过程的调控。本研究旨在对比Rag2敲除（Rag2 KO）小鼠与野生型同品系小鼠的脾脏微小RNA差异表达模式。
本研究从3只Rag2敲除小鼠与3只野生型小鼠体内分离获取脾脏组织，以此制备RNA样本，采用Affymetrix Genechip miRNA 4.0芯片开展微小RNA表达谱分析。该芯片在保留前代芯片高性能的基础上完成了内容更新，其覆盖范围全面，可靶向检测miRBase Release 20数据库中所有成熟微小RNA序列；配套分析文件包含宿主基因ID、预测与验证后的微小RNA靶基因以及成簇微小RNA相关信息，便于开展微小RNA结果的关联分析。
微小RNA表达芯片分析结果显示，Rag2敲除小鼠与野生型小鼠脾脏内存在大量差异表达的微小RNA。此类差异表达的微小RNA可能参与包括免疫调控在内的多种生物学与生理过程。
本研究结论表明，微小RNA可能在免疫调控过程中发挥活跃作用。
实验整体设计：采用Affymetrix Genechip miRNA 4.0芯片平台，对Rag2敲除小鼠与野生型小鼠的脾脏微小RNA进行表达谱分析，每组样本设置3次生物学重复。