title: Blocking of targeted microRNAs from next-generation sequencing libraries.
abstract: Highly abundant microRNAs (miRNAs) in small RNA sequencing libraries make it difficult to obtain efficient measurements of more lowly expressed species. We present a new method that allows for the selective blocking of specific, abundant miRNAs during preparation of sequencing libraries. This technique is specific with little off-target effects and has no impact on the reproducibility of the measurement of non-targeted species. In human plasma samples, we demonstrate that blocking of highly abundant hsa-miR-16-5p leads to improved detection of lowly expressed miRNAs and more precise measurement of differential expression overall. Furthermore, we establish the ability to target a second abundant miRNA and to multiplex the blocking of two miRNAs simultaneously. For small RNA sequencing, this technique could fill a similar role as do ribosomal or globin removal technologies in messenger RNA sequencing.
authors.affiliation: HudsonAlpha Institute for Biotechnology, Huntsville, AL 35806, USA.
authors.forename: Brian S
authors.initials: BS
authors.lastname: Roberts
authors.affiliation: HudsonAlpha Institute for Biotechnology, Huntsville, AL 35806, USA Department of Genetics, University of Alabama at Birmingham, Birmingham, AL 35294, USA.
authors.forename: Andrew A
authors.initials: AA
authors.lastname: Hardigan
authors.affiliation: HudsonAlpha Institute for Biotechnology, Huntsville, AL 35806, USA.
authors.forename: Marie K
authors.initials: MK
authors.lastname: Kirby
authors.affiliation: Center for Clinical and Translational Science, University of Alabama at Birmingham, Birmingham, AL 35294, USA.
authors.forename: Meredith B
authors.initials: MB
authors.lastname: Fitz-Gerald
authors.affiliation: Department of Medicine, Division of Gastroenterology and Hepatology, University of Alabama at Birmingham, Birmingham, AL 35294, USA.
authors.forename: C Mel
authors.initials: CM
authors.lastname: Wilcox
authors.affiliation: Center for Clinical and Translational Science, University of Alabama at Birmingham, Birmingham, AL 35294, USA Department of Medicine, Division of Clinical Immunology and Rheumatology, University of Alabama at Birmingham, Birmingham, AL 35294, USA.
authors.forename: Robert P
authors.initials: RP
authors.lastname: Kimberly
authors.affiliation: HudsonAlpha Institute for Biotechnology, Huntsville, AL 35806, USA rmyers@hudsonalpha.org.
authors.forename: Richard M
authors.initials: RM
authors.lastname: Myers
mesh_terms: D015723:Gene Library; D059014:High-Throughput Nucleotide Sequencing / Q000379:methods*; D006801:Humans; D035683:MicroRNAs / Q000097:blood* / Q000737:chemistry; D017423:Sequence Analysis, RNA / Q000379:methods*
chemical_list: C505362:MIRN16 microRNA, human; D035683:MicroRNAs