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[蜕膜胎盘细胞转录组分析方案]

[Protocol of Transcriptome Analysis of Decidual Placenta Cells].

作者信息

Babovskaya A A, Trifonova E A, Serebrova V N, Svarovskaya M G, Zarubin A A, Zhilyakova O V, Gabidulina T V, Poltanova A A, Rychkova L V, Stepanov V A

机构信息

Research Institute of Medical Genetics, Tomsk National Research Medical Center, Tomsk, 634050 Russia.

Siberian State Medical University, Tomsk, 634050 Russia.

出版信息

Mol Biol (Mosk). 2022 Mar-Apr;56(2):325-333. doi: 10.31857/S0026898422020045.

Abstract

The advent of high-throughput sequencing technologies has expanded our understanding of the biological significance of non-coding regions of the genome. In recent years, more and more studies have been devoted to studying the role of noncoding RNAs in the development of diseases, as well as their participation in various cellular processes. Until now, all transcriptome studies of native placental tissue with the description of the noncoding RNA region were carried out without isolating individual cell populations. This approach, due to the high cellular heterogeneity of the placental tissue, significantly complicates the ability to determine the molecular-biological functions of individual cells and their role in the molecular pathogenesis of reproductive disorders. In this work, we propose a technique for obtaining total RNA from single decidual cells of frozen placental tissue obtained by laser-capture microdissection technology for transcriptome sequencing, including a cluster of noncoding RNAs. This technique can be successfully used to study the full-genome expression profile of other placental cell populations. The high accuracy of results on the transcriptome profiling of decidual cells obtained using the developed technique was additionally confirmed by an integrative analysis with the results of a 10x Genomics experiment.

摘要

高通量测序技术的出现扩展了我们对基因组非编码区生物学意义的理解。近年来,越来越多的研究致力于研究非编码RNA在疾病发展中的作用,以及它们参与各种细胞过程的情况。到目前为止,所有对胎盘组织进行非编码RNA区域描述的转录组研究都是在未分离单个细胞群体的情况下进行的。由于胎盘组织细胞异质性高,这种方法极大地复杂化了确定单个细胞分子生物学功能及其在生殖障碍分子发病机制中作用的能力。在这项工作中,我们提出了一种从通过激光捕获显微切割技术获得的冷冻胎盘组织的单个蜕膜细胞中获取总RNA的技术,用于转录组测序,包括一组非编码RNA。该技术可成功用于研究其他胎盘细胞群体的全基因组表达谱。通过与10x基因组学实验结果的综合分析,进一步证实了使用所开发技术获得的蜕膜细胞转录组分析结果的高度准确性。

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