Polyethylene Glycol -Mediated Exosome Isolation: A Method for Exosomal RNA Analysis.

Q2 Biochemistry, Genetics and Molecular Biology Iranian Biomedical Journal Pub Date : 2024-03-01 DOI:10.61186/ibj.4129

Abdulwahab Teflischi Gharavi, Azadeh Niknejad, Saeed Irian, Amirabbas Rahimi, Mona Salimi

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Abstract

Background: : Exosomal RNAs (ExoRNAs) offer valuable insights into their cellular origin. ExoRNA studies were faced with challenges in obtaining sufficient amounts of high-quality RNA. Herein, we aimed to compare three traditional exosome isolation methods to introduce an appropriate strategy to extract RNA from cancer-derived exosomes for further RNA analysis.

Methods: Exosomes were isolated through ultracentrifugation, precipitation kit, and size exclusion column chromatography, and then characterized by dynamic light scattering and transmission electron microscopy, followed by extracting total RNA. The quality and quantity of the extracted RNAs were assessed by a NanoDrop and 2.5% agarose gel electrophoresis.

Results: Extracted exosomes displayed a similar range of size and morphology. We found that polyethylene glycol-precipitation method resulted in a higher RNA yield with a 260/280 ratio of 1.9. The obtained exoRNA appeared as a smear in the agarose gel, indicative of small exoRNAs.

Conclusion: We provide researchers a suitable approach to isolate exosomes based on yield and purity of exoRNA.

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聚乙二醇介导的外泌体分离：外泌体 RNA 分析方法。

背景：ExoRNA 为研究细胞起源提供了宝贵的线索。外泌体RNA研究面临着获取足量高质量RNA的挑战。在此，我们旨在比较三种传统的外泌体分离方法，以引入一种合适的策略从癌症衍生的外泌体中提取 RNA，用于进一步的 RNA 分析：方法：通过超速离心、沉淀试剂盒和尺寸排阻柱层析分离外泌体，然后用 DLS 和 TEM 对其进行表征，接着提取总 RNA。提取的 RNA 的质量和数量由 NanoDrop 和 2.5% 琼脂糖凝胶电泳进行评估：结果：提取的外泌体大小和形态相似。我们发现，PEG沉淀法的RNA产量更高，260/280比值为1.9。获得的外泌体 RNA 在琼脂糖凝胶中呈斑点状，表明是小的外泌体 RNA：我们为研究人员提供了一种根据外泌体RNA的产量和纯度分离外泌体的合适方法。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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