Homologous recombination Flashcards

1
Q

What is HR?

A

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

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2
Q

What cells are used in HR?

A

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

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3
Q

Describe gene knockouts

A

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

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4
Q

Describe gene knockins

A

Insert and expression of a gene

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5
Q

Describe a positive selection marker

A

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

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6
Q

Describe a negative selection marker

A

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

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7
Q

What are the 2 ways of transfecting cells?

A

Electroporation

Lipofection

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8
Q

Describe electroporation

A

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

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9
Q

Describe lipofection

A

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

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