Final Report Summary - E-DNA-T-PEP (Engineering DNA transfer into Cells by Precision in Electroporation)
The safe and effective delivery of DNA into target cells is the key issue in treating genetic and acquired diseases (such as cancer) by gene therapy and DNA vaccination. Delivery of naked DNA (gene) into cell is the safer alternative to viral vectors. Electroporation has been come a popular technique to permeabilize the membrane for different purposes such as medical treatments, food processing and biomass processing to deliver naked DNA into the living cells. Due to our lack of information about the fundamentals of electro-pores formation and DNA electro-transfer, electroporation method still suffers from low transfection efficiency, random uptake and excessive cell damage.
We seek to understand and control the transport of DNA in electroporation process at the molecular/sub-cellular level such that more efficient and safer non-viral gene delivery can be achieved. The introduction of naked DNA into living cell via non-viral routes is the safest approach in gene therapy. Furthermore, to understand the process of DNA electro-transfer, we investigated how DNA interacts in the confined environment (such as electro-pores), and moves through them using novel techniques such as nano-fluidics. To unveil the entire electroporation process, innovatively our group seeks to employed integrated single molecule techniques with micro/nano-fluidics to visualize the evolution of pore size and density at the membrane level (in high spatiotemporal resolutions). To this end we applied a multidisciplinary approach, combining disciplines as physical chemistry, transport phenomena, soft matter, DNA dynamics, biophysics and cell biology.
These advanced single macules techniques of DNA imaging integrated with micro/nano-fluidics enabled our laboratory to pioneer electroporation research at both fundamental and application level. We already investigated an exact energy landscape of the electropore formation and DNA electrotransfer into single cell. This research results in several important outcomes. First, it will help to develop electroporation methods to deliver naked DNA/gene into cell with high efficiency and minimal toxicity/cell damage for gene transfection and DNA vaccinations. Second, it will make it possible to optimize electroporation parameters beforehand, is crucial for efficient DNA/gene delivery. The successful outcome of this project will significantly aid the development of delivery of naked DNA into living cells, which will lead to electroporation-based therapies in the near future. In summary, the work in this project is an important step towards a detailed understanding of the DNA dynamics and transport into living cells.
In summary, the work in this project is an important step towards a detailed understanding of the electroporation of cellular membranes and DNA dynamics during electroporation, regarding the physics and the (bio-) chemistry of the DNA-membrane interaction. We are applying the obtained knowledge for the non-viral DNA delivery than can be employed in various fields. The obtained insight can be used to optimize these physical approaches for drug/gene delivery.
We seek to understand and control the transport of DNA in electroporation process at the molecular/sub-cellular level such that more efficient and safer non-viral gene delivery can be achieved. The introduction of naked DNA into living cell via non-viral routes is the safest approach in gene therapy. Furthermore, to understand the process of DNA electro-transfer, we investigated how DNA interacts in the confined environment (such as electro-pores), and moves through them using novel techniques such as nano-fluidics. To unveil the entire electroporation process, innovatively our group seeks to employed integrated single molecule techniques with micro/nano-fluidics to visualize the evolution of pore size and density at the membrane level (in high spatiotemporal resolutions). To this end we applied a multidisciplinary approach, combining disciplines as physical chemistry, transport phenomena, soft matter, DNA dynamics, biophysics and cell biology.
These advanced single macules techniques of DNA imaging integrated with micro/nano-fluidics enabled our laboratory to pioneer electroporation research at both fundamental and application level. We already investigated an exact energy landscape of the electropore formation and DNA electrotransfer into single cell. This research results in several important outcomes. First, it will help to develop electroporation methods to deliver naked DNA/gene into cell with high efficiency and minimal toxicity/cell damage for gene transfection and DNA vaccinations. Second, it will make it possible to optimize electroporation parameters beforehand, is crucial for efficient DNA/gene delivery. The successful outcome of this project will significantly aid the development of delivery of naked DNA into living cells, which will lead to electroporation-based therapies in the near future. In summary, the work in this project is an important step towards a detailed understanding of the DNA dynamics and transport into living cells.
In summary, the work in this project is an important step towards a detailed understanding of the electroporation of cellular membranes and DNA dynamics during electroporation, regarding the physics and the (bio-) chemistry of the DNA-membrane interaction. We are applying the obtained knowledge for the non-viral DNA delivery than can be employed in various fields. The obtained insight can be used to optimize these physical approaches for drug/gene delivery.