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Which one of the thermal approaches (heating DNA or cells) enhances the gene expression in mammalian cells?

OBJECTIVES: Heat treatment as a physical method could increase the cellular uptake of nucleic acids. In this study, the effects of heat shock were evaluated to enhance the transfection efficiency of three plasmid DNAs into HeLa and TC-1 cancerous, and HEK-293 T and Vero non-cancerous cell lines usin...

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Autores principales: Milani, Alireza, Bolhassani, Azam, Rouhollah, Fatemeh, Naseroleslami, Maryam
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Springer Netherlands 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8418791/
https://www.ncbi.nlm.nih.gov/pubmed/34482511
http://dx.doi.org/10.1007/s10529-021-03176-0
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author Milani, Alireza
Bolhassani, Azam
Rouhollah, Fatemeh
Naseroleslami, Maryam
author_facet Milani, Alireza
Bolhassani, Azam
Rouhollah, Fatemeh
Naseroleslami, Maryam
author_sort Milani, Alireza
collection PubMed
description OBJECTIVES: Heat treatment as a physical method could increase the cellular uptake of nucleic acids. In this study, the effects of heat shock were evaluated to enhance the transfection efficiency of three plasmid DNAs into HeLa and TC-1 cancerous, and HEK-293 T and Vero non-cancerous cell lines using lipofectamine 2000 reagent. METHODS: Two methods of cell- and DNA-based heat treatment were used. Heating DNA solution was performed at 94 °C for 5, 10 and 15 min, and also 72 °C for 30, 60 and 120 min, individually. Moreover, heating the cells was done by incubation at 42 °C for 2 h in different times such as before, during and after DNA transfection. RESULTS: Our data showed that the conformation of plasmid DNAs was changed at different temperatures with increasing time. The heat-treated plasmid DNAs (94 °C for 10 min or 72 °C for 30 min) indicated higher transfection efficiency than untreated plasmid DNAs (p < 0.05). Furthermore, heat treatment of cells before and during the transfection was higher than untreated cells (p < 0.01). Our results demonstrated that DNA transfection efficiency in cancerous cells was less than non-cancerous cells (p < 0.01). CONCLUSION: Generally, these findings showed that transfection mediated by thermal stimulation could enhance gene transfection in mammalian cell lines.
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spelling pubmed-84187912021-09-07 Which one of the thermal approaches (heating DNA or cells) enhances the gene expression in mammalian cells? Milani, Alireza Bolhassani, Azam Rouhollah, Fatemeh Naseroleslami, Maryam Biotechnol Lett Original Research Paper OBJECTIVES: Heat treatment as a physical method could increase the cellular uptake of nucleic acids. In this study, the effects of heat shock were evaluated to enhance the transfection efficiency of three plasmid DNAs into HeLa and TC-1 cancerous, and HEK-293 T and Vero non-cancerous cell lines using lipofectamine 2000 reagent. METHODS: Two methods of cell- and DNA-based heat treatment were used. Heating DNA solution was performed at 94 °C for 5, 10 and 15 min, and also 72 °C for 30, 60 and 120 min, individually. Moreover, heating the cells was done by incubation at 42 °C for 2 h in different times such as before, during and after DNA transfection. RESULTS: Our data showed that the conformation of plasmid DNAs was changed at different temperatures with increasing time. The heat-treated plasmid DNAs (94 °C for 10 min or 72 °C for 30 min) indicated higher transfection efficiency than untreated plasmid DNAs (p < 0.05). Furthermore, heat treatment of cells before and during the transfection was higher than untreated cells (p < 0.01). Our results demonstrated that DNA transfection efficiency in cancerous cells was less than non-cancerous cells (p < 0.01). CONCLUSION: Generally, these findings showed that transfection mediated by thermal stimulation could enhance gene transfection in mammalian cell lines. Springer Netherlands 2021-09-05 2021 /pmc/articles/PMC8418791/ /pubmed/34482511 http://dx.doi.org/10.1007/s10529-021-03176-0 Text en © The Author(s), under exclusive licence to Springer Nature B.V. 2021 This article is made available via the PMC Open Access Subset for unrestricted research re-use and secondary analysis in any form or by any means with acknowledgement of the original source. These permissions are granted for the duration of the World Health Organization (WHO) declaration of COVID-19 as a global pandemic.
spellingShingle Original Research Paper
Milani, Alireza
Bolhassani, Azam
Rouhollah, Fatemeh
Naseroleslami, Maryam
Which one of the thermal approaches (heating DNA or cells) enhances the gene expression in mammalian cells?
title Which one of the thermal approaches (heating DNA or cells) enhances the gene expression in mammalian cells?
title_full Which one of the thermal approaches (heating DNA or cells) enhances the gene expression in mammalian cells?
title_fullStr Which one of the thermal approaches (heating DNA or cells) enhances the gene expression in mammalian cells?
title_full_unstemmed Which one of the thermal approaches (heating DNA or cells) enhances the gene expression in mammalian cells?
title_short Which one of the thermal approaches (heating DNA or cells) enhances the gene expression in mammalian cells?
title_sort which one of the thermal approaches (heating dna or cells) enhances the gene expression in mammalian cells?
topic Original Research Paper
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8418791/
https://www.ncbi.nlm.nih.gov/pubmed/34482511
http://dx.doi.org/10.1007/s10529-021-03176-0
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