Spontaneous cellular uptake of exogenous messenger RNA in vivo is nucleic acid-specific, saturable and ion dependent
Abstract
The development of new treatments in the post-genomic era requires methods for safe delivery of foreign genetic information in vivo. As a transient, natural and controllable alternative to recombinant viruses or plasmid DNA (pDNA), purified or in vitro transcribed messenger RNA (mRNA) can be used for the expression of any therapeutic protein in vitro and in vivo. As it has been shown previously, the simple injection of naked mRNA results in local uptake and expression. We show here that this process, in the skin, can greatly be modulated according to the injection solution composition and blocked by an excess of competing nucleic acids or a drug affecting cytosolic mobility. Different cell types at the site of injection can take up the foreign nucleic acid molecules and the protein translated from this is detected for no more than a few days. To test this gene transfer method in humans, we produced in vitro transcribed mRNA under good manufacturing practice (GMP) conditions in a dedicated facility. After injection into the human dermis, we could document the translation of the exogenous mRNA. Our results pave the way toward the use of mRNA as a vehicle for transient gene delivery in humans.
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The lack of biological efficacy of HPLC-purified basic Epo mRNA highlights the importance and potency of sequence-engineering.
“Although high performance liquid chromatography (HPLC) purification depletes contaminants and, thus, contributes to reduced immunostimulation as well as improves mRNA expression to some extent,36,37,38 the lack of biological efficacy of HPLC-purified basic Epo mRNA highlights the importance and potency of sequence-engineering.”
From Sequence-engineered mRNA without chemical nucleoside modifications enables an effective protein therapy in large animals (2015) · cites this paper as [37]
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- Authors
- Probst, J., Weide, B., Scheel, B., Pichler, B. J., Hoerr, I., Rammensee, H.-G., & Pascolo, S.
- Affiliations
- University of Tübingen (DE)
- CureVac (Germany) (DE)
- Corresponding author
- Pascolo, S
- Venue
- Gene Therapy
- Published
- 2007-05-03
- Volume
- 14
- Issue
- 15
- Pages
- 1175-1180
- ISSN
- 0969-7128 (ISSN-L)
- 1476-5462
- OpenAlex
- W1991064719
- Cited by
- 206
- References
- 17
- Citations by year
- 2026: 4 · 2025: 14 · 2024: 13 · 2023: 22 · 2022: 18 · 2021: 36 · 2020: 11 · 2019: 10 · 2018: 11 · 2017: 4 · 2016: 8 · 2015: 10 · 2014: 8 · 2013: 5 · 2012: 6
- FWCI
- 2.25
- Type
- journal article
- Language
- en
- Update notices
- erratum (2009-05-14)
- Open access
- closed
- Locations
- Gene Therapy (publishedVersion)
- PubMed (publishedVersion)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA Interference and Gene Delivery
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA and protein synthesis mechanisms
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA Research and Splicing
- Keywords
- mRNA delivery, skin, dermis, ion dependence, in vitro transcribed mRNA, mRNA translation, therapeutic protein expression
- MeSH terms
- Animals, DNA, Humans, Hydrogen-Ion Concentration, Immunohistochemistry, Injections, Subcutaneous, Luciferases, Nucleic Acids, RNA, Messenger, Skin, Transfection, Genetic Therapy, Gene Expression, Microscopy, Confocal, Mice
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