Hello everyone
I am transfecting my primary endothelial cells using Lipofectamine LTX. As controls, besides other controls such as non-transfected cells and the transfected cells with empty plasmid, I included Lipofectamine-only control (or control with no DNA) in my experiment. 48h post-transfection, I detected an increase in the expression level of my gene of interest in Lipofectamine-only control compared to data obtained from 24h post-transfection samples. I am wondering if the cell stress induced by lipofectamine in my cells could affect the expression of my gene of interest.
If anyone have any experience in this issue, I would appreciate if they could share their experience or knowledge with me.
Yes, Lipofectamine alone can influence gene expression, particularly due to cellular stress responses. Studies have shown that liposome-based transfection reagents, including Lipofectamine, can induce stress-related gene expression changes. This effect is likely due to membrane perturbation, oxidative stress, and activation of stress-response pathways, such as heat shock proteins (HSPs).
In your case, the increase in gene expression at 48 hours post-transfection could be due to:
- Cellular stress response triggered by Lipofectamine, leading to upregulation of stress-related genes.
- Inflammatory or metabolic changes induced by lipid-based transfection reagents.
- Delayed secondary effects where cells adapt to the presence of Lipofectamine over time.
To confirm whether Lipofectamine is directly affecting your gene of interest, you could:
Please check this reference for more details:
Article Liposome-based DNA carriers may induce cellular stress respo...
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