Depletion of erythropoietic miR-486-5p and miR-451a improves detectability of rare microRNAs in peripheral blood-derived small RNA sequencing libraries
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Abstract
Erythroid-specific miR-451a and miR-486-5p are two of the most dominant microRNAs (miRNAs) in human peripheral blood. In small RNA sequencing libraries, their overabundance reduces diversity as well as complexity and consequently causes negative effects such as missing detectability and inaccurate quantification of low abundant miRNAs. Here we present a simple, cost-effective and easy to implement hybridization-based method to deplete these two erythropoietic miRNAs from blood-derived RNA samples. By utilization of blocking oligonucleotides, this method provides a highly efficient and specific depletion of miR-486-5p and miR-451a, which leads to a considerable increase of measured expression as well as detectability of low abundant miRNA species. The blocking oligos are compatible with common 5′ ligation-dependent small RNA library preparation protocols, including commercially available kits, such as Illumina TruSeq and Perkin Elmer NEXTflex. Furthermore, the here described method and oligo design principle can be easily adapted to target many other miRNA molecules, depending on context and research question.
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- Authors
- Juzenas, S., Lindqvist, C. M., Ito, G., Dolshanskaya, Y., Halfvarson, J., Franke, A., & Hemmrich-Stanisak, G.
- Affiliations
- Institute of Clinical Molecular Biology, Christian-Albrechts-University of Kiel , DE 24105 Kiel, Germany
- Örebro University (SE)
- Christian-Albrechts-Universität zu Kiel (DE)
- Venue
- NAR Genomics and Bioinformatics
- Publisher
- Oxford University Press (OUP)
- Published
- 2020-02-12
- Volume
- 2
- Issue
- 1
- Pages
- lqaa008-lqaa008
- ISSN
- 2631-9268 (ISSN-L)
- PMC
- PMC7671325
- OpenAlex
- W3006049475
- Cited by
- 17
- References
- 38
- Citations by year
- 2026: 2 · 2025: 3 · 2024: 2 · 2023: 1 · 2022: 3 · 2021: 4 · 2020: 2
- FWCI
- 0.80
- Type
- journal article
- Language
- en
- Open access
- gold
- doi.org
- Licence
- Locations
- NAR Genomics and Bioinformatics (publishedVersion, cc-by-nc)
- bioRxiv (Cold Spring Harbor Laboratory) (acceptedVersion, cc-by-nc)
- PubMed Central (publishedVersion, cc-by-nc)
- PubMed (publishedVersion)
- Lithuanian University of Health Sciences (submittedVersion)
- Funders
- Deutsche Forschungsgemeinschaft (EXC2167)
- Swedish Foundation for Strategic Research (RB13-0160, BIO IBD)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Cancer Research › MicroRNA in disease regulation
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Gene expression and cancer classification
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Genomics and Phylogenetic Studies
- Keywords
- miR-486-5p, miR-451a
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