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Single cell quantification of microRNA from small numbers of non-invasively sampled primary human cells

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Title: Single cell quantification of microRNA from small numbers of non-invasively sampled primary human cells
Authors: Ho, V
Baker, J
Willison, K
Barnes, P
Donnelly, L
Klug, D
Item Type: Journal Article
Abstract: Expression levels of microRNAs (miRNAs) in single cells are low and conventional miRNA detection methods require amplification that can be complex, time-consuming, costly and may bias results. Single cell microfluidic platforms have been developed; however, current approaches are unable to absolutely quantify single miRNA molecules expressed in single cells. Herein, we present an amplification-free sandwich hybridisation assay to detect single miRNA molecules in single cells using a microfluidic platform that optically traps and lyses individual cells. Absolute quantification of miR-21 and miR-34a molecules was achieved at a single cell level in human cell lines and validated using real-time qPCR. The sensitivity of the assay was demonstrated by quantifying single miRNA molecules in nasal epithelial cells and CD3+ T-cells, as well as nasal fluid collected non-invasively from healthy individuals. This platform requires ~50 cells or ~30 µL biofluid and can be extended for other miRNA targets therefore it could monitor miRNA levels in disease progression or clinical studies.
Issue Date: 26-Apr-2023
Date of Acceptance: 17-Apr-2023
URI: http://hdl.handle.net/10044/1/104073
DOI: 10.1038/s42003-023-04845-8
ISSN: 2399-3642
Publisher: Nature Portfolio
Start Page: 1
End Page: 11
Journal / Book Title: Communications Biology
Volume: 6
Copyright Statement: © The Author(s) 2023 Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/ licenses/by/4.0/.
Publication Status: Published
Article Number: 458
Online Publication Date: 2023-04-26
Appears in Collections:Chemistry
Biological and Biophysical Chemistry
National Heart and Lung Institute
Faculty of Medicine
Faculty of Natural Sciences

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