Homologous recombination

Question 1

What is HR?

Answer 1

A single copy of the modified gene is modified in its normal chromosomal environment and allows for the natural repair mechanism of non-dividing cells to add the transgene

Question 2

What cells are used in HR?

Answer 2

Embryonic stem cells
From the inner cell mass of the blastocyst
Can develop into any type of cell

Question 3

Describe gene knockouts

Answer 3

A gene is inserted and expresses a marker gene in a position that results in a natural gene being inactivated

Question 4

Describe gene knockins

Answer 4

Insert and expression of a gene

Question 5

Describe a positive selection marker

Answer 5

The presence of a gene in the sequence that provides resistance and therefore survives

Question 6

Describe a negative selection marker

Answer 6

The presence of a gene in the sequence results in cell death

Question 7

What are the 2 ways of transfecting cells?

Answer 7

Electroporation

Lipofection

Question 8

Describe electroporation

Answer 8

Expose cells to an electric field which briefly opens the pores in the cell membrane and allows for the DNA to pass-through
This can damage the cells

Question 9

Describe lipofection

Answer 9

Cationic lipids form liposome vesicles with DNA and then bind with the cell membrane