title: Evaluation of inhibition of miRNA expression induced by anti-miRNA oligonucleotides.
abstract: MicroRNAs (miRNAs) are short RNA molecules that control the expression of mRNAs associated with various biological processes. Therefore, deregulated miRNAs play an important role in the pathogenesis of diseases. Numerous studies aimed at developing novel miRNA-based drugs or determining miRNA functions have been conducted by inhibiting miRNAs using anti-miRNA oligonucleotides (AMOs), which inhibit the function by hybridizing with miRNA. To increase the binding affinity and specificity to target miRNA, AMOs with various chemical modifications have been developed. Evaluating the potency of these various types of AMOs is an essential step in their development. In this study, we developed a capillary electrophoresis with laser-induced fluorescence (CE-LIF) method to evaluate the potency of AMOs by measuring changes in miRNA levels with fluorescence-labeled ssDNA probes using AMO-miR-23a, which inhibits miR-23a related to lung cancer. In order to eliminate interference by excess AMOs during hybridization of the ssDNA probe with the miR-23a, the concentration of the ssDNA probe was optimized. This newly developed method was used to compare the potency of two different modified AMOs. The data were supported by the results of a luciferase assay. This study demonstrated that CE-LIF analysis could be used to accurately evaluate AMO potency in biological samples.
authors.affiliation: Molecular Recognition Research Center, Korea Institute of Science and Technology, Hwarangno 14-gil 5, Seongbuk-gu, Seoul, 02792, South Korea.|College of Life Sciences and Biotechnology, Korea University, Seoul, 02841, South Korea.
authors.forename: Dong-Kyu
authors.initials: DK
authors.lastname: Chae
authors.affiliation: Molecular Recognition Research Center, Korea Institute of Science and Technology, Hwarangno 14-gil 5, Seongbuk-gu, Seoul, 02792, South Korea.
authors.forename: Eunmi
authors.initials: E
authors.lastname: Ban
authors.affiliation: Molecular Recognition Research Center, Korea Institute of Science and Technology, Hwarangno 14-gil 5, Seongbuk-gu, Seoul, 02792, South Korea.
authors.forename: Young Sook
authors.initials: YS
authors.lastname: Yoo
authors.affiliation: College of Life Sciences and Biotechnology, Korea University, Seoul, 02841, South Korea.
authors.forename: Ja-Hyun
authors.initials: JH
authors.lastname: Baik
authors.affiliation: Molecular Recognition Research Center, Korea Institute of Science and Technology, Hwarangno 14-gil 5, Seongbuk-gu, Seoul, 02792, South Korea. ejsong@kist.re.kr.
authors.forename: Eun Joo
authors.initials: EJ
authors.lastname: Song
mesh_terms: D000070416:Antagomirs / Q000008:administration & dosage* / Q000235:genetics*; D015536:Down-Regulation / Q000187:drug effects / Q000235:genetics*; D019075:Electrophoresis, Capillary / Q000379:methods*; D005786:Gene Expression Regulation / Q000187:drug effects / Q000235:genetics*; D035683:MicroRNAs / Q000096:biosynthesis* / Q000235:genetics; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013050:Spectrometry, Fluorescence / Q000379:methods*
chemical_list: D000070416:Antagomirs; D035683:MicroRNAs
keywords: Anti-miRNA oligonucleotide; Capillary electrophoresis with laser-induced fluorescence; miR-23a; miRNA