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bioRxiv · 10.1101/2025.01.10.632138

MiRNA Locker: A Modularized DNA Assembly As miRNA Inhibitors

Zhu, L.·Min, L.·Zhu, C.-s.·Wu, X.·Li, W.·Ma, J.·Lei, Y.·Gao, C.·Qiu, X.·Liu, C.
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Abstract

MicroRNAs (miRNAs) are vital post-transcriptional regulators that govern key cellular processes such as proliferation, migration, and apoptosis. Current loss-of-function approaches, including chemically modified antisense oligonucleotides (ASOs), face significant challenges, including high costs, limited scalability, and off-target effects. To overcome these limitations, we developed "miRNA Locker", a novel miRNA inhibition platform created using the Overlapped Oligo Assembly (OOA) method. This innovative platform constructs highly stable dumbbell-shaped single-stranded DNA structures, offering improved target specificity, scalability, and cost-effectiveness. Using miR-214 as a proof-of-concept target, we demonstrated that miRNA Lockers effectively bind Argonaute-miRNA complexes, reduce miRNA levels, and induce downstream changes in gene expression and cellular phenotypes, surpassing the performance of commercial antagomirs. Furthermore, applying miRNA Lockers to miR-654 validated the regulatory role of miR-654 in modulating RNF8 expression and promoting epithelial-mesenchymal transition (EMT) in lung cancer cells. Our results highlight the potential of miRNA Lockers as a versatile tool for studying miRNA function and advancing miRNA-based therapies. Graphical Abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=114 SRC="FIGDIR/small/632138v1_ufig1.gif" ALT="Figure 1"> View larger version (26K): org.highwire.dtl.DTLVardef@c6ac1eorg.highwire.dtl.DTLVardef@14e2698org.highwire.dtl.DTLVardef@1c7d84aorg.highwire.dtl.DTLVardef@148f9c1_HPS_FORMAT_FIGEXP M_FIG C_FIG

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Zhu, L., Min, L., Zhu, C.-s., Wu, X., Li, W., Ma, J., Lei, Y., Gao, C., Qiu, X., Liu, C.. 2025-01-11. MiRNA Locker: A Modularized DNA Assembly As miRNA Inhibitors. https://doi.org/10.1101/2025.01.10.632138

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