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Efficient expression of stabilized mRNA PEG-peptide polyplexes in liver

Gene Therapy volume 22, pages 993–999 (2015)Cite this article

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Abstract

The expression efficiency in liver following hydrodynamic delivery of in vitro transcribed mRNA was improved 2000-fold using a codon-optimized mRNA luciferase construct with flanking 3' and 5' human β-globin untranslated regions (UTR mRNA) over an unoptimized mRNA without β-globin UTRs. Nanoparticle UTR mRNA polyplexes were formed using a novel polyacridine polyethylene glycol (PEG) peptide, resulting in an additional 15-fold increase in expression efficiency in the liver. The combined increase in expression for UTR mRNA PEG-peptide polyplexes was 3500-fold over mRNA lacking UTRs and PEG-peptide. The expression efficiency of UTR mRNA polyplex was 10-fold greater than the expression from an equivalent 1 μg dose of pGL3. Maximal expression was maintained from 4 to 24 h. Serum incubation established the unique ability of the polyacridine PEG-peptide to protect UTR mRNA polyplexes from RNase metabolism by binding to double-stranded regions. UTR mRNA PEG-peptide polyplexes are efficient nonviral vectors that circumvent the need for a nuclear uptake, representing an advancement toward the development of a targeted gene delivery system to transfect liver hepatocytes.

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Acknowledgements

We gratefully acknowledge support from NIH Grants GM097093 and T32 GM008365 (STC and JAP).

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  1. Division of Medicinal and Natural Products Chemistry, College of Pharmacy, University of Iowa, Iowa City, IA, USA

    S T Crowley, J A Poliskey, N J Baumhover & K G Rice

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  1. S T Crowley
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Correspondence to K G Rice.

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Crowley, S., Poliskey, J., Baumhover, N. et al. Efficient expression of stabilized mRNA PEG-peptide polyplexes in liver. Gene Ther 22, 993–999 (2015). https://doi.org/10.1038/gt.2015.68

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