Abstract
Small non-coding RNAs (ncRNAs), less than 200 nucleotides in length, play important roles in various biological processes, such as pre-mRNA splicing, pre-rRNA processing and modification, and gene expression regulation. However, characterization of small ncRNAs remains difficult mainly due to methodological obstacles in selective reduction of these RNAs. Here we describe an approach to deplete small ncRNAs, in principle any types of RNAs, using second generation antisense oligonucleotide-directed RNase H cleavage pathway in human cells. This protocol includes oligonucleotide design, transfection, RNA preparation, and target RNA detection.
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Acknowledgement
This work was funded by an internal funding from ISIS Pharmaceuticals.
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Liang, XH., Shen, W., Crooke, S.T. (2015). Efficient and Selective Knockdown of Small Non-Coding RNAs. In: Rederstorff, M. (eds) Small Non-Coding RNAs. Methods in Molecular Biology, vol 1296. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-2547-6_19
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DOI: https://doi.org/10.1007/978-1-4939-2547-6_19
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-2546-9
Online ISBN: 978-1-4939-2547-6
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