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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...
Autores principales: | , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
MDPI
2015
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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. |
format | Online Article Text |
id | pubmed-4588190 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2015 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
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 |
work_keys_str_mv | AT uchidasatoshi screeningofmrnachemicalmodificationtomaximizeproteinexpressionwithreducedimmunogenicity AT kataokakazunori screeningofmrnachemicalmodificationtomaximizeproteinexpressionwithreducedimmunogenicity AT itakakeiji screeningofmrnachemicalmodificationtomaximizeproteinexpressionwithreducedimmunogenicity |