title: Efficient expression of stabilized mRNA PEG-peptide polyplexes in liver.
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.
authors.affiliation: Division of Medicinal and Natural Products Chemistry, College of Pharmacy, University of Iowa, Iowa City, IA, USA.
authors.forename: S T
authors.initials: ST
authors.lastname: Crowley
authors.affiliation: Division of Medicinal and Natural Products Chemistry, College of Pharmacy, University of Iowa, Iowa City, IA, USA.
authors.forename: J A
authors.initials: JA
authors.lastname: Poliskey
authors.affiliation: Division of Medicinal and Natural Products Chemistry, College of Pharmacy, University of Iowa, Iowa City, IA, USA.
authors.forename: N J
authors.initials: NJ
authors.lastname: Baumhover
authors.affiliation: Division of Medicinal and Natural Products Chemistry, College of Pharmacy, University of Iowa, Iowa City, IA, USA.
authors.forename: K G
authors.initials: KG
authors.lastname: Rice
mesh_terms: D000818:Animals; D004247:DNA / Q000235:genetics / Q000378:metabolism; D015870:Gene Expression; D018014:Gene Transfer Techniques; D005822:Genetic Vectors; D006801:Humans; D008099:Liver / Q000378:metabolism / Q000502:physiology*; D051379:Mice; D010455:Peptides / Q000235:genetics* / Q000378:metabolism*; D010957:Plasmids / Q000235:genetics / Q000378:metabolism; D011092:Polyethylene Glycols / Q000378:metabolism*; D020871:RNA Stability / Q000502:physiology; D012333:RNA, Messenger / Q000096:biosynthesis* / Q000235:genetics / Q000378:metabolism; D014018:Tissue Distribution; D014158:Transcription, Genetic; D014162:Transfection / Q000379:methods*; D020506:Untranslated Regions / Q000235:genetics; D055544:beta-Globins / Q000235:genetics
chemical_list: D010455:Peptides; D012333:RNA, Messenger; D020506:Untranslated Regions; D055544:beta-Globins; D011092:Polyethylene Glycols; D004247:DNA