mRNA Transfection-Induced Activation of Primary Human Monocytes and Macrophages: Dependence on Carrier System and Nucleotide Modification.

Scientific Reports
Hanieh MoradianManfred Gossen

Abstract

Monocytes and macrophages are key players in maintaining immune homeostasis. Identifying strategies to manipulate their functions via gene delivery is thus of great interest for immunological research and biomedical applications. We set out to establish conditions for mRNA transfection in hard-to-transfect primary human monocytes and monocyte-derived macrophages due to the great potential of gene expression from in vitro transcribed mRNA for modulating cell phenotypes. mRNA doses, nucleotide modifications, and different carriers were systematically explored in order to optimize high mRNA transfer rates while minimizing cell stress and immune activation. We selected three commercially available mRNA transfection reagents including liposome and polymer-based formulations, covering different application spectra. Our results demonstrate that liposomal reagents can particularly combine high gene transfer rates with only moderate immune cell activation. For the latter, use of specific nucleotide modifications proved essential. In addition to improving efficacy of gene transfer, our findings address discrete aspects of innate immune activation using cytokine and surface marker expression, as well as cell viability as key readouts to j...Continue Reading

References

Nov 4, 2000·International Journal of Pharmaceutics·S DokkaY Rojanasakul
Jun 18, 2003·Trends in Immunology·David M SansomYong Zheng
Jan 5, 2005·Biochemical and Biophysical Research Communications·Tim J L Van De ParreGuido R Y De Meyer
Feb 25, 2005·Molecular Therapy : the Journal of the American Society of Gene Therapy·Abdelatif Elouahabi, Jean-Marie Ruysschaert
Oct 10, 2007·Methods in Enzymology·Ewa Grudzien-NogalskaEdward Darzynkiewicz
Dec 29, 2007·The Journal of Pathology·X Zhang, D M Mosser
Apr 7, 2009·Journal of Immunological Methods·Michael SchnoorStefan Lorkowski
Jan 19, 2010·International Journal of Pharmaceutics·S ZouJ G Hecker
Apr 3, 2010·European Journal of Pharmaceutical Sciences : Official Journal of the European Federation for Pharmaceutical Sciences·Conchita Tros de IlarduyaNejat Düzgüneş
Feb 16, 2012·Molecular Therapy : the Journal of the American Society of Gene Therapy·Katalin KarikóDrew Weissman
Jan 12, 2013·Journal of Controlled Release : Official Journal of the Controlled Release Society·Kyle K L PhuaSmita K Nair
Nov 26, 2013·Environmental Toxicology and Chemistry·Katja KettlerA Jan Hendriks
Feb 11, 2014·Methods in Molecular Biology·Isabel SiegertJonathan Jantsch
May 24, 2014·Nature Reviews. Immunology·Florent Ginhoux, Steffen Jung
Jun 4, 2014·Immunology and Cell Biology·Anthony TroegelerGeanncarlo Lugo-Villarino
Jun 26, 2014·Cellular Immunology·Andrew J MitchellWolfgang Weninger
Sep 17, 2014·Journal of Visualized Experiments : JoVE·Marten B MaeßStefan Lorkowski
Sep 18, 2014·Clinical Hemorheology and Microcirculation·Toralf RochAndreas Lendlein
Sep 23, 2014·Nature Reviews. Drug Discovery·Ugur SahinÖzlem Türeci
Feb 11, 2015·Analytical Biochemistry·Marten B MaeßStefan Lorkowski
Apr 12, 2015·Annual Review of Immunology·Chen VarolSteffen Jung
Apr 22, 2015·Journal of Immunological Methods·Olga SchererStefan Lorkowski
Jul 1, 2015·Expert Opinion on Biological Therapy·Hyewon Youn, June-Key Chung
Sep 1, 2015·Molecular Therapy : the Journal of the American Society of Gene Therapy·Drew Weissman, Katalin Karikó
Oct 8, 2015·Clinical Hemorheology and Microcirculation·Thanga Bhuvanesh Vijaya BhaskarToralf Roch
Jan 1, 2016·Journal of Controlled Release : Official Journal of the Controlled Release Society·Kevin J KauffmanDaniel G Anderson
May 28, 2016·Experimental Biology and Medicine·Molly E OgleEdward A Botchwey
Jul 28, 2016·Biochemistry. Biokhimii︠a︡·O V SergeevaT S Zatsepin
Oct 22, 2016·Arteriosclerosis, Thrombosis, and Vascular Biology·Rachel M KratofilJustin F Deniset
Nov 5, 2016·Annual Review of Physiology·Peter J Murray
Feb 12, 2017·Methods in Molecular Biology·Charles A Warwick, Yuriy M Usachev
Oct 31, 2017·Methods : a Companion to Methods in Enzymology·Sanders Oh, John A Kessler
Apr 25, 2018·Nucleic Acid Therapeutics·Matthew G Stanton
May 16, 2018·Methods in Molecular Biology·Andrea-Anneliese KellerStefan Lorkowski
Sep 10, 2018·Molecular Therapy. Nucleic Acids·Sriram VaidyanathanAnton P McCaffrey

❮ Previous
Next ❯

Citations

Nov 5, 2020·Journal of Molecular Medicine : Official Organ of the Gesellschaft Deutscher Naturforscher Und Ärzte·Hanieh MoradianManfred Gossen
May 26, 2021·Advanced Drug Delivery Reviews·Elias Fattal, François Fay
Jun 17, 2021·Journal of Materials Chemistry. B, Materials for Biology and Medicine·Feng DingChuanxu Yang
Jul 3, 2021·Viruses·Natalia Teresa JarzebskaSteve Pascolo

❮ Previous
Next ❯

Methods Mentioned

BETA
transfections
transfection
in
in vitro transcription
flow cytometry
flow
transfect
electrophoreses
electrophoresis
density gradient centrifugation

Software Mentioned

Elements
NIS
FlowJo
GraphPad Prism

Related Concepts

Trending Feeds

COVID-19

Coronaviruses encompass a large family of viruses that cause the common cold as well as more serious diseases, such as the ongoing outbreak of coronavirus disease 2019 (COVID-19; formally known as 2019-nCoV). Coronaviruses can spread from animals to humans; symptoms include fever, cough, shortness of breath, and breathing difficulties; in more severe cases, infection can lead to death. This feed covers recent research on COVID-19.

Blastomycosis

Blastomycosis fungal infections spread through inhaling Blastomyces dermatitidis spores. Discover the latest research on blastomycosis fungal infections here.

Nuclear Pore Complex in ALS/FTD

Alterations in nucleocytoplasmic transport, controlled by the nuclear pore complex, may be involved in the pathomechanism underlying multiple neurodegenerative diseases including Amyotrophic Lateral Sclerosis and Frontotemporal Dementia. Here is the latest research on the nuclear pore complex in ALS and FTD.

Applications of Molecular Barcoding

The concept of molecular barcoding is that each original DNA or RNA molecule is attached to a unique sequence barcode. Sequence reads having different barcodes represent different original molecules, while sequence reads having the same barcode are results of PCR duplication from one original molecule. Discover the latest research on molecular barcoding here.

Chronic Fatigue Syndrome

Chronic fatigue syndrome is a disease characterized by unexplained disabling fatigue; the pathology of which is incompletely understood. Discover the latest research on chronic fatigue syndrome here.

Evolution of Pluripotency

Pluripotency refers to the ability of a cell to develop into three primary germ cell layers of the embryo. This feed focuses on the mechanisms that underlie the evolution of pluripotency. Here is the latest research.

Position Effect Variegation

Position Effect Variagation occurs when a gene is inactivated due to its positioning near heterochromatic regions within a chromosome. Discover the latest research on Position Effect Variagation here.

STING Receptor Agonists

Stimulator of IFN genes (STING) are a group of transmembrane proteins that are involved in the induction of type I interferon that is important in the innate immune response. The stimulation of STING has been an active area of research in the treatment of cancer and infectious diseases. Here is the latest research on STING receptor agonists.

Microbicide

Microbicides are products that can be applied to vaginal or rectal mucosal surfaces with the goal of preventing, or at least significantly reducing, the transmission of sexually transmitted infections. Here is the latest research on microbicides.