International Journal of Applied Science and Engineering
Published by Chaoyang University of Technology

Retno Lestari 1*, Bismi Yasinta Maharani 1,2, Damai Ria Setyawati 2, Etik Mardliyati 2, Ankan Dutta Chowdhury 3, Tarwadi 2,4*

1 Department of Biology, Faculty of Mathematics and Natural Sciences, Universitas Indonesia, West Java, 16424 Indonesia

2 Research Centre for Vaccine and Drug, National Research and Innovation Agency (BRIN), Banten, 15314 Indonesia

3 Amity Institute of Nanotechnology, Amity University Kolkata, Kolkata, West Bengal 700135, India

4 PT Indomabs Biosantika Utama, Gedung Technology Business and Innovation Centre (TBIC), Kabupaten Bogor, West Java, 16340 Indonesia

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ABSTRACT

Branched low molecular weight PEI (bPEI-800 Da) is capable of condensing DNA molecules effectively. However, it has poor capacity to facilitate gene delivery. In this research, bPEI-800 is formulated with cholesterol, and Tween 20 or Tween 80 to form a liposome in order to obtain an effective transfection agent. The liposome's capacity to condense DNA was assessed using an ethidium bromide exclusion assay. Meanwhile, the ability of the liposome to protect DNA from DNase was evaluated by a DNA mobility shift assay and an enzymatic degradation assay using DNA gel electrophoresis. Transmission electronic microscopy (TEM) analysis was performed to determine the liposome/DNA particle size. Cell viability and transgene expression of the green fluorescence protein (GFP)-encoding gene were evaluated in HEK-293T cells. It was revealed that the bPEI-800-based liposome is able to condense DNA into nanoparticles 50–100 nm and protect DNA molecules from DNase I degradation. Furthermore, the bPEI-800-based liposome enhanced the transgene expression up to 1.6-fold, as a higher number of GFP-expressing cells were detected compared to bPEI-800 alone under the fluorescence microscope. Interestingly, the presence of cholesterol in the liposome formulation also reduced cytotoxicity, as revealed in MTT assay data. In summary, the bPEI-800-based liposome allowed condensation of DNA molecules into nanoparticle sizes, and enhanced transgene expression in HEK-293T cells. Further exploration in formulating and evaluating the bPEI-800-based liposome in other mammalian cells is needed to achieve a safe and effective transfection agent.


Keywords: Gene expression, Liposome, Polyethyleneimine, Transfection agent.


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ARTICLE INFORMATION

Received: 2023-10-26
Revised: 2023-11-30
Accepted: 2023-12-11


Cite this article:

Lestari, R., Maharani, B.Y., Setyawati, D.R., Mardliyati, E., Chowdhury, A.D., Tarwadi 2024. Enhanced transgene expression of branched polyethyleneimine (bPEI-800)-based liposome nanoparticles in HEK-293T cells. International Journal of Applied Science and Engineering, 21, 2023445. https://doi.org/10.6703/IJASE.202409_21(4).003

  Copyright The Author(s). This is an open access article distributed under the terms of the Creative Commons Attribution License (CC BY 4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are cited.

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