高通量测序、生物芯片、qPCR等是研究miRNA的主要方法。前者可以发现新的microRNA，而后两者这要是研究不同实验组的已知microRNA的差异表达。以下文献（Schotte D et al Leukemia 2011，25, 1389-1399）即是对7种不同基因型的小儿急性淋巴细胞白血病样本进行深度测序以及生物信息学分析预测，以期发现新的、ALL特异的miRNA。最终，该研究确定了554 个已知miRNA, 28新的miRNA以及 431 可能的新的miRNA，其中53 个已知miRNA, 16 个新的miRNA以及170 个可能的新miRNA为ALL特异表达。该研究扩充了我们对ALL相关miRNA及可能靶基因的了解，并为深度测序发现疾病相关的新miRNA提供方法借鉴。
Discovery of new microRNAs by small RNAome deep sequencing in childhood acute lymphoblastic leukemia
Abstract MicroRNAs (miRNAs) relevant to acute lymphoblastic leukemia (ALL) in children are hypothesized to be largely unknown as most miRNAs have been identified in non-leukemic tissues.In order to discover these miRNAs, we applied high-throughput sequencing to pooled fractions of leukemic cells obtained from 89 pediatric cases covering seven well-defined genetic types of ALL and normal hematopoietic cells. This resulted into 78 million small RNA reads representing 554 known, 28 novel and 431 candidate novel miR genes. In all, 153 known, 16 novel and 170 candidate novel mature miRNAs and miRNA-star strandswere only expressed in ALL, whereas 140 known, 2 novel and 82 candidate novel mature miRNAs and miRNA-star strands were unique to normal hematopoietic cells. Stem-loop reverse transcriptase (RT)-quantitative PCR analyses confirmed the differential expression of selected mature miRNAs in ALL types and normal cells. Expression of 14 new miRNAs inversely correlated with expression of predicted target genes (_0.49p Spearman's correlation coefficients (Rs)p_0.27, Pp0.05); among others, low levels of novel sol-miR-23 associated with high levels of its predicted (antiapoptotic) target BCL2 (B-cell lymphoma 2) in precursor B-ALL (Rs _0.36, P?0.007). The identification of 41000 miR genes expressed in different types of ALL forms a comprehensive repository for further functional studies that address the role of miRNAs in the biology of ALL.
原文链接地址：http://www.nature.com/leu/journal/v25/n9/full/leu2011105a.html
