Brief Heat Shock Increases Stable Integration of Lipid-Mediated DNA Transfections

Pipes, Brian L.; Vasanwala, Farha H.; Tsang, Tom C.; Zhang, Tong; Luo, Phoebe; Harris, David T.

doi:10.2144/05381bm05

Cited by 9 publications

(4 citation statements)

References 10 publications

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“…Although the temperature increase on the surface of the magnetic pulse generator could also have contributed to enhanced transfection as reported previously ( 27 ), it should be noted however, that this increase was most significant in cells transfected with superparamagnetic particles and thus can be attributed to nanoparticle properties in the presence of the pulsating magnetic field. The alternating magnetic field, used in our study, was at lower frequencies and thus any possibility that the superparamagnetic particles are producing heat when exposed to higher frequencies ( 28 ) can be ruled out.…”

Section: Discussionmentioning

confidence: 52%

Enhancement of the efficiency of non-viral gene delivery by application of pulsed magnetic field

Kamau

Hassa²,

Steitz³

et al. 2006

Nucleic Acids Research

111

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New approaches to increase the efficiency of non-viral gene delivery are still required. Here we report a simple approach that enhances gene delivery using permanent and pulsating magnetic fields. DNA plasmids and novel DNA fragments (PCR products) containing sequence encoding for green fluorescent protein were coupled to polyethylenimine coated superparamagnetic nanoparticles (SPIONs). The complexes were added to cells that were subsequently exposed to permanent and pulsating magnetic fields. Presence of these magnetic fields significantly increased the transfection efficiency 40 times more than in cells not exposed to the magnetic field. The transfection efficiency was highest when the nanoparticles were sedimented on the permanent magnet before the application of the pulsating field, both for small (50 nm) and large (200–250 nm) nanoparticles. The highly efficient gene transfer already within 5 min shows that this technique is a powerful tool for future in vivo studies, where rapid gene delivery is required before systemic clearance or filtration of the gene vectors occurs.

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Section: Discussionmentioning

confidence: 52%

Enhancement of the efficiency of non-viral gene delivery by application of pulsed magnetic field

Kamau

Hassa²,

Steitz³

et al. 2006

Nucleic Acids Research

111

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show abstract

“…change in fluidity of the cell membrane), and/ or an increased stable integration rate ( i.e. change in fluidity of the nuclear membrane or in chromatin structure) (Pipes et al 2005 ). In our study, heat treatment of the cells at 42 °C for 2 h before transfection could increase transfection efficiency and gene expression in both non-cancerous ( p < 0.01) and cancerous ( p < 0.05) cell lines.…”

Section: Discussionmentioning

confidence: 99%

“…These DNA modifications may influence the transfection efficiency (Hou et al 2008 ). Previous in vitro experiments showed that heat treatment of the cells increased lipid-mediated DNA transfection efficiency (Pipes et al 2005 ; Takizaki et al 2017 ) through caveolar endocytosis, and subsequently escaping from lysosomal digestion (Takizaki et al 2017 ).…”

Section: Introductionmentioning

confidence: 99%

Which one of the thermal approaches (heating DNA or cells) enhances the gene expression in mammalian cells?

et al. 2021

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Objectives Heat treatment as a physical method could increase the cellular uptake of nucleic acids. In this study, the effects of heat shock were evaluated to enhance the transfection efficiency of three plasmid DNAs into HeLa and TC-1 cancerous, and HEK-293 T and Vero non-cancerous cell lines using lipofectamine 2000 reagent. Methods Two methods of cell- and DNA-based heat treatment were used. Heating DNA solution was performed at 94 °C for 5, 10 and 15 min, and also 72 °C for 30, 60 and 120 min, individually. Moreover, heating the cells was done by incubation at 42 °C for 2 h in different times such as before, during and after DNA transfection. Results Our data showed that the conformation of plasmid DNAs was changed at different temperatures with increasing time. The heat-treated plasmid DNAs (94 °C for 10 min or 72 °C for 30 min) indicated higher transfection efficiency than untreated plasmid DNAs ( p < 0.05). Furthermore, heat treatment of cells before and during the transfection was higher than untreated cells ( p < 0.01). Our results demonstrated that DNA transfection efficiency in cancerous cells was less than non-cancerous cells ( p < 0.01). Conclusion Generally, these findings showed that transfection mediated by thermal stimulation could enhance gene transfection in mammalian cell lines.

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“…Cationic lipids interact with anionic DNA and the conjugates are used for gene delivery into cells 5 , 6 . Brief heat shock enhanced the transfection efficiency, but the effects was moderate 7 . Lipofectamine 2000 reagent is currently and widely used for gene delivery procedure 8 , however its transfection rate depends on cell types and cell toxicity of the reagent is a crucial issue.…”

Section: Introductionmentioning

confidence: 94%

Caspase inhibition improves viability and efficiency of liposomal transfection

Yoshida,

Yamasaki,

Tadagaki

2023

Sci Rep

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High transfection efficiency is the most important point for experiments of DNA and RNA introduction into cells. Decrease of cell viability during the transfection procedure is a crucial issue, resulting in transfection failure. However, the mechanism underlying cell growth inhibition has not been fully elucidated. Lipofection is frequently used for transfection experiments, whereases, depending on cell type, it causes a decrease in cell viability. The present study demonstrates here that a potent pan-caspase inhibitor Q-VD-OPh blocked cell death during the lipofection, indicating apoptosis was induced in lipofection. Moreover, Q-VD-OPh drastically increased transfected cells. This method provides easier and more effective transfection system of lipofection and may be useful for transfection of not only cell lines but also clinical uses such as gene therapy and nucleic acids vaccine.

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Brief Heat Shock Increases Stable Integration of Lipid-Mediated DNA Transfections

Cited by 9 publications

References 10 publications

Enhancement of the efficiency of non-viral gene delivery by application of pulsed magnetic field

Enhancement of the efficiency of non-viral gene delivery by application of pulsed magnetic field

Which one of the thermal approaches (heating DNA or cells) enhances the gene expression in mammalian cells?

Caspase inhibition improves viability and efficiency of liposomal transfection

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