Impact of physicochemical properties of DNA/PEI complexes on transient transfection of mammalian cells pdf

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21-07-2022
Social Studies

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Impact of physicochemical properties of DNA/PEI complexes on transient transfection of mammalian cells pdf

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tutorconsortium346 tutorconsortium346 · Answer 1 · 2022-08-02T19:46:03+02:00

Transient gene expression (TGE) has been successfully accomplished in mammalian cell cultures using polyethyleneimine (PEI). The connection between the synthesis of DNA/PEI complexes and their biological activity has not yet been conclusively proven.

What are properties of DNA?

DNA, also known as deoxyribonucleic acid, is the genetic substance found in practically all prokaryotes and eukaryotes. The cell's nucleus is where DNA is found, however mitochondrial DNA, a small amount of DNA, is also present there. As a result, your body's circulating cells all contain the same kind of DNA. The body of living things depends on DNA in many ways. Here are a few justifications for the importance of DNA.
Every cell's genetic material is carried by DNA, which also faithfully duplicates itself throughout cell division.
Body structure, messengers, enzymes, and hormones are all formed in large part by the interaction between DNA and proteins.
Rarely, DNA undergoes alteration, creating mutations that result in genetic differences and evolution.

What is the impact of physicochemical properties of DNA?

A virus-like particle (VLP) production platform was used to conduct a systematic investigation on DNA/PEI complexes, their physicochemical characteristics during formation, and their transfection effectiveness.
Instead of using simple ionic solutions, the same chemically specified cell culture medium for DNA/PEI complex formation was used to reduce changes in complex characteristics during transfection.
An initial concentration of 1E + 10 DNA/PEI complexes/mL partially aggregated after creation, with an average size of 300 nm. By using X-ray spectroscopy, the role of NaCl ions in the evolution of complexes was examined, highlighting the importance of complexing media composition in TGE techniques.
At the moment of transfection, it was estimated that each cell had 250 complexes plus aggregates after 15 minutes of incubation. Such heterogeneous preparations are difficult to define, thus cryo-electron microscopy and nanoparticle tracking analysis (NTA) were coupled to get a full view of the preparation.
Drug blocking endocytosis was used to assess the contribution of each DNA/PEI complex subpopulation. Intriguingly, all complexes effectively transported DNA, and when high size aggregates—which enter through micropinocytosis—were suppressed, they significantly increased the effectiveness of transfection.

The physicochemical components involved in DNA delivery procedures must be understood. As a result, this study offers fresh perspectives on the characterization of DNA/PEI complexes that will help with more successful and dependable TGE methods.

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