Optimization of cationic polymer-mediated transfection for RNA interference

Transfection efficiency was estimated to optimize the conditions for RNA interference (RNAi), including transfection time, validity, and nucleic acid concentration and type, using the EZ Trans Cell Reagent, a cationic polymer. An shRNA against GFP was designed and transfected into cells using the EZ...

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Autores principales: Fan, Xiaojie, Yang, Jingnan, Wu, Guangyao, Wang, Meiyi, Cheng, Xiaoxia, Liu, Chang, Liu, Qian, Wen, Yanan, Meng, Shuangshuang, Wang, Zhenxing, Lin, Xuhong, An, Lei
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Sociedade Brasileira de Genética 2022
Materias:
Cellular, Molecular and Developmental Genetics
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8915406/
https://www.ncbi.nlm.nih.gov/pubmed/35275159
http://dx.doi.org/10.1590/1678-4685-GMB-2021-0237
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author Fan, Xiaojie
Yang, Jingnan
Wu, Guangyao
Wang, Meiyi
Cheng, Xiaoxia
Liu, Chang
Liu, Qian
Wen, Yanan
Meng, Shuangshuang
Wang, Zhenxing
Lin, Xuhong
An, Lei
author_facet Fan, Xiaojie
Yang, Jingnan
Wu, Guangyao
Wang, Meiyi
Cheng, Xiaoxia
Liu, Chang
Liu, Qian
Wen, Yanan
Meng, Shuangshuang
Wang, Zhenxing
Lin, Xuhong
An, Lei
author_sort Fan, Xiaojie
collection PubMed
description Transfection efficiency was estimated to optimize the conditions for RNA interference (RNAi), including transfection time, validity, and nucleic acid concentration and type, using the EZ Trans Cell Reagent, a cationic polymer. An shRNA against GFP was designed and transfected into cells using the EZ transfection reagent. The shRNA significantly decreased the expression of GFP. In addition, pre-diluted transfection reagent at room temperature and small nucleic acids increased the transfection efficiency, which peaked at 24 h. Compared with circular nucleic acids, linear nucleic acids showed higher transfection efficiency and a higher genome integration rate. We optimized cationic polymer-mediated RNAi conditions, and our data will be useful for future RNAi studies.
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spelling pubmed-89154062022-03-29 Optimization of cationic polymer-mediated transfection for RNA interference Fan, Xiaojie Yang, Jingnan Wu, Guangyao Wang, Meiyi Cheng, Xiaoxia Liu, Chang Liu, Qian Wen, Yanan Meng, Shuangshuang Wang, Zhenxing Lin, Xuhong An, Lei Genet Mol Biol Cellular, Molecular and Developmental Genetics Transfection efficiency was estimated to optimize the conditions for RNA interference (RNAi), including transfection time, validity, and nucleic acid concentration and type, using the EZ Trans Cell Reagent, a cationic polymer. An shRNA against GFP was designed and transfected into cells using the EZ transfection reagent. The shRNA significantly decreased the expression of GFP. In addition, pre-diluted transfection reagent at room temperature and small nucleic acids increased the transfection efficiency, which peaked at 24 h. Compared with circular nucleic acids, linear nucleic acids showed higher transfection efficiency and a higher genome integration rate. We optimized cationic polymer-mediated RNAi conditions, and our data will be useful for future RNAi studies. Sociedade Brasileira de Genética 2022-03-09 /pmc/articles/PMC8915406/ /pubmed/35275159 http://dx.doi.org/10.1590/1678-4685-GMB-2021-0237 Text en https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution License
spellingShingle Cellular, Molecular and Developmental Genetics
Fan, Xiaojie
Yang, Jingnan
Wu, Guangyao
Wang, Meiyi
Cheng, Xiaoxia
Liu, Chang
Liu, Qian
Wen, Yanan
Meng, Shuangshuang
Wang, Zhenxing
Lin, Xuhong
An, Lei
Optimization of cationic polymer-mediated transfection for RNA interference
title Optimization of cationic polymer-mediated transfection for RNA interference
title_full Optimization of cationic polymer-mediated transfection for RNA interference
title_fullStr Optimization of cationic polymer-mediated transfection for RNA interference
title_full_unstemmed Optimization of cationic polymer-mediated transfection for RNA interference
title_short Optimization of cationic polymer-mediated transfection for RNA interference
title_sort optimization of cationic polymer-mediated transfection for rna interference
topic Cellular, Molecular and Developmental Genetics
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8915406/
https://www.ncbi.nlm.nih.gov/pubmed/35275159
http://dx.doi.org/10.1590/1678-4685-GMB-2021-0237
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