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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Detalles Bibliográficos
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
Descripción
Sumario: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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