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Gene delivery into Siberian sturgeon cell lines by commercial transfection reagents

  • Ji Hun Lee
  • Seung Tae Lee
  • Yoon Kwon Nam
  • Seung Pyo GongEmail author
Article
  • 19 Downloads

Abstract

The optimal transfection conditions for efficient transgene delivery into a specific cell type should be empirically determined, particularly in cases involving unusual cell types. We compared the conditions for effective introduction of transgenes into Siberian sturgeon (Acipenser baerii) cell lines by evaluating the cytotoxicity and transfection efficiency of three commercially available transfection reagents: Lipofectamine 2000, X-tremeGENE HP DNA Transfection Reagent, and GeneJuice Transfection Reagent. Plasmid vectors containing the gene encoding enhanced green fluorescent protein were mixed with each of the transfection reagents using reagent-to-plasmid ratios of 1:1, 2:1, and 4:1. Then, the complexes were used to transfect three Siberian sturgeon cell lines derived from the heart, head kidney, and gonad. Cytotoxicity and transfection efficiency were measured via flow cytometry after propidium iodide staining. No significant cytotoxicity was observed at the optimal treatment conditions in all cases, with the exception of Lipofectamine 2000-treated gonad-derived cells. Although the transfection efficiencies in A. baerii cells were generally low, X-tremeGENE HP DNA Transfection Reagent showed the highest transfection efficiency at ratios of 2:1 or 4:1, depending on the cell type. Hence, X-tremeGENE HP DNA Transfection Reagent can be used to effectively transfer foreign genes into three A. baerii cell lines.

Keywords

Gene delivery Siberian sturgeon Cell lines Transfection reagent 

Notes

Funding information

This research was a part of the project (20170327) funded by Korea Institute of Marine Science & Technology Promotion (KIMST), the Ministry of Oceans and Fisheries, Korea.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict in interest.

Supplementary material

11626_2018_316_MOESM1_ESM.docx (113 kb)
ESM 1 (DOCX 113 kb)

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Copyright information

© The Society for In Vitro Biology 2018

Authors and Affiliations

  1. 1.Department of Fisheries BiologyPukyong National UniversityBusanSouth Korea
  2. 2.Department of Animal Life ScienceKangwon National UniversityChuncheonSouth Korea
  3. 3.Department of Marine-Biomaterials and AquacultureCollege of Fisheries Science, Pukyong National UniversityBusanSouth Korea

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