Isolation and characterization of microRNAs of human mature erythrocytes.
Isolation and characterization of microRNAs of human mature erythrocytes.
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DOI：
10.1007/978-1-60761-811-9_13
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发表时间：
2010
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人成熟红细胞是终末分化的细胞，在发育过程中失去了它们的细胞核和细胞器。即使成熟的红细胞缺乏核糖体和其他大尺寸的RNA，它们仍然保留小尺寸的RNA。我们最近通过使用几种不同的技术，包括北方印迹、miRNA微阵列和实时PCR，发现成熟红细胞中存在丰富且多样的microRNA。此外，分级分离和基因组分析已经揭示，红细胞microRNA表达与网织红细胞或白细胞的microRNA表达不同，并且成熟红细胞贡献了全血中的大部分microRNA表达。因此，循环红细胞中microRNA表达的全球分析有可能为红细胞生物学和红细胞相关疾病提供机制见解。在此，我们提供了从人成熟红细胞中分离和表征microRNA的详细方法，以使此类研究能够用于涉及红细胞的人类疾病。
Human mature erythrocytes are terminally differentiated cells that have lost their nuclei and organelles during development. Even though mature erythrocytes lack ribosomal and other large-sized RNAs, they still retain small-sized RNAs. We have recently shown that there are abundant and diverse species of microRNAs in mature erythrocytes through the use of several different techniques, including northern blot, miRNA microarray, and real-time PCR. Furthermore, fractionation and genomic analysis has revealed that erythrocyte microRNA expression is different from that of reticulocytes or leukocytes and that mature erythrocytes contribute the majority of microRNA expression in whole blood. Therefore, global analysis of microRNA expression in circulating erythrocytes has the potential to provide mechanistic insights into erythrocyte biology and erythrocyte-related disorders. Here, we have provided the detailed methods for isolating and characterizing the microRNAs from human mature erythrocytes to enable such researches into human diseases involving erythrocytes.