Transfection of DNA into Eukaryotes

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Transfection of DNA into Eukaryotes

• Mom, please, may I have a glowing fish, too?

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Although glowing fish are fun, the procedure used
to make them is very useful to scientists!
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We are going to make living cells glow
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Transfection

• Delivery of foreign molecules (DNA) into
  eukaryotic cells
• Cell begins to read the piece of DNA and produce
  the protein encoded on it

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How can we get DNA into a cell?

• Inject it
• Poke holes in the membrane without destroying the
  cells
• Create a complex of DNA plus some substance that
  will go into the cell

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We will use Cationic liposomes

• It creates an artificial membrane
• Membrane formed around pieces of DNA
• Cell membrane will fuse with this artificial
  membrane and bring the DNA into the cell

Liposomes are also be useful for drug and
cosmetic delivery
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• Liposome-DNA complex taken up by the cell by
  fusing with the cell membrane

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• Can change an entire animal or just a culture of
  cells
• View with fluorescent microscope
• Use blue or ultraviolet light

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GFP

• Green fluorescent protein
• Gives luminescent jelly fish their greenish glow

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Plasmid Vector

• GFP is a marker protein on a plasmid vector
• A plasmid is a piece of DNA coding for a specific
  protein
• If the cell read the DNA and produces the
  protein, it will also produce GFP.
• That particular protein will glow
• If there is enough of that protein, the entire
  cell will glow

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What can we do with glowing cells?

• Determine distribution of protein
• Track movement of protein
• Track movement of cells
• VERY useful for scientists!