Absolute quantification of mammalian microRNAs for therapeutic RNA cleavage and detargeting
- Carolyn Kraus1,6,
- Jiayi Wang1,4,6,
- Haiying Zheng2,
- Jennifer Broderick1,
- Nandagopal Ajaykumar2,
- Mina Zamani2,
- Mengqi Yang1,
- Katharine Cecchini1,
- Shun-qing Liang3,
- Olena Kolumba2,
- Kathryn Chase1,
- Jooyoung Lee1,5,
- Wen Xue1,
- Erik J. Sontheimer1 and
- Ildar Gainetdinov2
- 1RNA Therapeutics Institute, University of Massachusetts Chan Medical School, Worcester, Massachusetts 01655, USA
- 2Department of Biology, New York University, New York, New York 10003, USA
- 3Division of Gastroenterology, Hepatology, and Nutrition, University of Minnesota Twin Cities, Minneapolis, Minnesota 55455, USA
- Corresponding author: ildar.gainetdinov{at}nyu.edu
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↵6 These authors contributed equally to this work.
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Handling editor: Mihaela Zavolan
Abstract
MicroRNAs (miRNAs) are small regulatory RNAs that destabilize partially complementary transcripts and cleave perfectly paired targets. miRNAs are often expressed in a specific tissue, allowing miRNA-directed cleavage to be used to prevent genome editing or gene replacement therapy in unintended cell types, a strategy called detargeting. miRNA intracellular concentration influences the potency of gene silencing, yet the absolute steady-state levels of just a few miRNAs have been determined in mammalian tissues. Here, we report the absolute abundance of miRNAs in 14 human and mouse cell lines and 17 mouse tissues, including eight brain regions. Our experiments in human cultured cells demonstrate that both miRNA and target levels influence efficacy of cleavage of fully complementary transcripts. We report the miRNA concentration required for productive cleavage of highly expressed transcripts and identify mouse miRNAs that reach this threshold in vivo.
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Footnotes
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Article is online at http://www.rnajournal.org/cgi/doi/10.1261/rna.080566.125.
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Freely available online through the RNA Open Access option.
- Received April 28, 2025.
- Accepted May 4, 2025.
This article, published in RNA, is available under a Creative Commons License (Attribution 4.0 International), as described at http://creativecommons.org/licenses/by/4.0/.
