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Regulation of miRNA Switches Depends on Nucleotide Modification, Number of Target Sites and Complementarity
EP27103
Poster Title: Regulation of miRNA Switches Depends on Nucleotide Modification, Number of Target Sites and Complementarity
Submitted on 08 Feb 2018
Author(s): John Lockhart, John Canfield, Ezinne Mong, Jeffrey VanWye, Hana Totary-Jain
Affiliations: Department of Molecular Pharmacology and Physiology, Morsani College of Medicine, University of South Florida, Tampa, Florida 33612
This poster was presented at USF Research Day 2018
Poster Views: 465
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Poster Information
Abstract: mRNA-based therapeutics represent a game-changing category of biological drugs that holds great promise for the treatment of human diseases1,2. While the incorporation of naturally occurring modified nucleotides during the synthesis of mRNA has greatly increased their potency and safety3, challenges in selective expression have hindered some clinical applications. Recently, microRNA-regulated in vitro transcribed mRNAs, called miRNA switches, have been used to successfully target the expression of exogenous mRNA in a cell-selective manner4. However, the effect of nucleotide modifications commonly used in miRNA switches on microRNA-dependent silencing has not been examined. Here we show that pseudouridine-modified and unmodified miRNA switches are equally silenced by microRNA, and the silencing is more effective with increasing numbers of fully complementary miRNA target sites. In contrast, the silencing of pseudouridine/5-methylcytosine-modified miRNA switches is not improved by additional microRNA target sites and is affected by the sequence of the microRNA target sites. Both pseudouridine-modified and pseudouridine/5-methylcytosine-modified miRNA switches with target sites complementary to only the microRNA seed sequence are less effectively silenced compared to unmodified miRNA switches. This work provides insights into the regulation of miRNA switches and can inform the design of cell-selective mRNA therapeutics.
Summary: mRNA therapeutics are an up-and-coming category of biologics. The ability to control mRNA therapeutics using endogenous regulatory systems (i.e. microRNA) greatly increases their specificity and safety. However, the effect of modifications used during the synthesis of mRNA therapeutics on the regulatory capacity of microRNAs has not been assessed. Here we show that these modifications can significantly alter the regulation dependent on microRNA sequence, target site number, and complementarity.References: Sahin U, Karikó K, Tureci O. mRNA-based therapeutics - developing a new class of drugs. Nat Rev Drug Discov. 13:759–780. [PMID: 25233993]

Huang CL, Leblond AL, Turner EC, Kumar AH, et al. Synthetic chemically modified mRNA-based delivery of cytoprotective factor promotes early cardiomyocyte survival post-acute myocardial infarction. Mol Pharm. 12:991–996. [PMID: 25588055]

Kormann MSD, Hasenpusch G, Aneja MK, Nica G, et al. Expression of therapeutic proteins after delivery of chemically modified mRNA in mice. Nat Biotechnol. 29:154–157. [PMID: 21217696]

Miki K, Endo K, Takahashi S, Funakoshi S, et al. Efficient Detection and Purification of Cell Populations Using Synthetic MicroRNA Switches. Cell Stem Cell. 16:699–711. [PMID: 26004781]

Wroblewska L, Kitada T, Endo K, Siciliano V, et al. Mammalian synthetic circuits with RNA binding proteins for RNA-only delivery. Nat Biotechnol. 33:839–41. [PMID: 26237515]





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