Researchers have a wide variety of gene delivery techniques to introduce plasmid DNA, siRNA or duplex RNAi, oligonucleotides, and RNA into eukaryotic cells for a variety of research and drug discovery applications. A review of these techniques is provided on the table below.
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Lipid delivery using cationic lipids
How it works:
DNA, siRNA, or oligonucleotides and transfection reagent
(commercially available) are diluted in separate tubes.




(commercially available) are diluted in separate tubes.

DNA and transfection reagent combined to form complexes.
Positive charge on cationic lipid binds to phosphate backbone on nucleic acid.
Positive charge on cationic lipid binds to phosphate backbone on nucleic acid.

Complexes added to cells. Positive charge on cationic lipid helps bind complex to membrane.

Complexes enter the cell via endocytosis.

Assay for gene expression or silencing.
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Viral delivery
How it works:
Generate recombinant virus via gene cloning, cell culture, virus isolation,
and clone characterization.




and clone characterization.

Prepare and purify high titer viral vectors.

Infect cells (containing viral receptor) at appropriate multiplicity of infection (MOI).

Remove virus and/or add fresh medium.

Assay for gene expression or silencing.
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Delivery by electroporation
How it works:
Prepare cells by suspending in electroporation buffer.
Pulse cells electrically in the presence of specialized buffer and DNA.



Pulse cells electrically in the presence of specialized buffer and DNA.

Electrical pulse creates a potential difference across the membrane and induces
temporary pores in the cell membrane for DNA entry
temporary pores in the cell membrane for DNA entry

Cells returned to growth conditions.

Assay for gene expression or silencing.
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Delivery by other chemical methods, i. e. calcium phosphate precipitation, DEAE-dextran, polybrene
How it works:
Prepare solutions. Mix calcium chloride (in phosphate buffer) and DNA.

Allow precipitation of extremely small, insoluble particles containing condensed DNA



Allow precipitation of extremely small, insoluble particles containing condensed DNA

CaPO
4: Add complexes to cell culture and incubate.
Complexes adhere to cell membranes, and enter into the cytoplasm by endocytosis.
Complexes adhere to cell membranes, and enter into the cytoplasm by endocytosis.

DEAE-Dextran and polybrene:
Complexes are delivered by osmotic shock using DMSO or glycerol.
Positive-charged polymers complex with negatively charged DNA molecules to enable binding to the cell surface.

Complexes are delivered by osmotic shock using DMSO or glycerol.
Positive-charged polymers complex with negatively charged DNA molecules to enable binding to the cell surface.

Remove complexes from cells, wash, and add fresh culture medium.

Assay for gene expression or silencing.

Assay for gene expression or silencing.
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For Research Use Only. Not for use in diagnostic procedures.