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Screening of mRNA Chemical Modification to Maximize Protein Expression with Reduced Immunogenicity

Chemical modification of nucleosides in mRNA is an important technology to regulate the immunogenicity of mRNA. In this study, various previously reported mRNA formulations were evaluated by analyzing in vitro protein expression and immunogenicity in multiple cell lines. For the macrophage-derived c...

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Detalles Bibliográficos
Autores principales: Uchida, Satoshi, Kataoka, Kazunori, Itaka, Keiji
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2015
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4588190/
https://www.ncbi.nlm.nih.gov/pubmed/26213960
http://dx.doi.org/10.3390/pharmaceutics7030137
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author Uchida, Satoshi
Kataoka, Kazunori
Itaka, Keiji
author_facet Uchida, Satoshi
Kataoka, Kazunori
Itaka, Keiji
author_sort Uchida, Satoshi
collection PubMed
description Chemical modification of nucleosides in mRNA is an important technology to regulate the immunogenicity of mRNA. In this study, various previously reported mRNA formulations were evaluated by analyzing in vitro protein expression and immunogenicity in multiple cell lines. For the macrophage-derived cell line, RAW 264.7, modified mRNA tended to have reduced immunogenicity and increased protein expression compared to the unmodified mRNA. In contrast, in some cell types, such as hepatocellular carcinoma cells (HuH-7) and mouse embryonic fibroblasts (MEFs), protein expression was decreased by mRNA modification. Further analyses revealed that mRNA modifications decreased translation efficiency but increased nuclease stability. Thus, mRNA modification is likely to exert both positive and negative effects on the efficiency of protein expression in transfected cells and optimal mRNA formulation should be determined based on target cell types and transfection purposes.
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spelling pubmed-45881902015-10-08 Screening of mRNA Chemical Modification to Maximize Protein Expression with Reduced Immunogenicity Uchida, Satoshi Kataoka, Kazunori Itaka, Keiji Pharmaceutics Article Chemical modification of nucleosides in mRNA is an important technology to regulate the immunogenicity of mRNA. In this study, various previously reported mRNA formulations were evaluated by analyzing in vitro protein expression and immunogenicity in multiple cell lines. For the macrophage-derived cell line, RAW 264.7, modified mRNA tended to have reduced immunogenicity and increased protein expression compared to the unmodified mRNA. In contrast, in some cell types, such as hepatocellular carcinoma cells (HuH-7) and mouse embryonic fibroblasts (MEFs), protein expression was decreased by mRNA modification. Further analyses revealed that mRNA modifications decreased translation efficiency but increased nuclease stability. Thus, mRNA modification is likely to exert both positive and negative effects on the efficiency of protein expression in transfected cells and optimal mRNA formulation should be determined based on target cell types and transfection purposes. MDPI 2015-07-23 /pmc/articles/PMC4588190/ /pubmed/26213960 http://dx.doi.org/10.3390/pharmaceutics7030137 Text en © 2015 by the authors; licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Uchida, Satoshi
Kataoka, Kazunori
Itaka, Keiji
Screening of mRNA Chemical Modification to Maximize Protein Expression with Reduced Immunogenicity
title Screening of mRNA Chemical Modification to Maximize Protein Expression with Reduced Immunogenicity
title_full Screening of mRNA Chemical Modification to Maximize Protein Expression with Reduced Immunogenicity
title_fullStr Screening of mRNA Chemical Modification to Maximize Protein Expression with Reduced Immunogenicity
title_full_unstemmed Screening of mRNA Chemical Modification to Maximize Protein Expression with Reduced Immunogenicity
title_short Screening of mRNA Chemical Modification to Maximize Protein Expression with Reduced Immunogenicity
title_sort screening of mrna chemical modification to maximize protein expression with reduced immunogenicity
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4588190/
https://www.ncbi.nlm.nih.gov/pubmed/26213960
http://dx.doi.org/10.3390/pharmaceutics7030137
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