Transformation of mammalian cells

Question 1

What type of mammalian cells do we transform?

Answer 1

Embryonic stem cells in a cell culture

Question 2

What are the 6 methods to transform mammalian cells?

Answer 2

Calcium phosphate polymers, optoperforation, liposome mediated, viral transduction, electroporation, microinjection

Question 3

Which 3 transformation methods are most commonly used to transform mammalian cells?

Answer 3

Calcium phosphate polymers, electroporation, liposome mediated

Question 4

How does transformation with calcium phosphate polymers work?

Answer 4

Mix DNA in a calcium phosphate solution and pipet it on top of cells, which will form a precipitate on the cell surface. The precipitate alters membrane permeability and the DNA gets into the cell

Question 5

How does transformation with optoperforation work?

Answer 5

Use a laser to poke holes in the membrane

Question 6

How does liposome mediated transformation work?

Answer 6

Mix DNA into a phospholipid solution, which will form vesicles/liposomes in solution and then fuse with the cell to release their DNA into the cell

Question 7

How does transformation with viral transduction work?

Answer 7

Use a disabled virus containing YFG as a delivery method, but the virus has to be engineered to do this

Question 8

How does transformation with electroporation work?

Answer 8

Exactly the same as with yeast. Uses electricity to poke holes in the membrane and the DNA gets in that way

Question 9

How does transformation with microinjection work?

Answer 9

Uses a very tiny needle to inject the DNA directly into the nucleus. Very difficult to do, you need to get the needle though two membranes and back out without damaging the cell