Cell Culture - Lecture 8: Gene Transfer at Hochschule Bonn-Rhein-Sieg

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What are the two types of transfection?

Transient transfection
Stable transfection

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For which type of transfection are plasmids used?

Transient

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What are barriers within the cell that may prevent transfer of exogenous DNA?

Lysosome/Endosome
Nucleases

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By which mechanism is exogenous DNA internalized into target cells?

Endocytosis

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Name three chemical procedures to introduce DNA into cells.

Calcium Phosphate
DEAE Dextran
Artificial liposomes

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For what reason are transfection reagents positively charged?

To bind to negatively charged DNA backbone

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What are three disadvantages about using Calcium Phosphate for transfection?

Crystals are toxic to transfected cells
Can cause mutation in transfected DNA
Not all cell lines can be transfected
(+ 1: Efficiency is not as high as lipofection)

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In the calcium phosphate transfection method, DNA is introduced into the cells as a ___ with calcium phosphate. The solid result can be observed under the microscope.

In the calcium phosphate transfection method, DNA is introduced into the cells as a co-precipitate with calcium phosphate. The solid result can be observed under the microscope.

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Why is the DEAE Dextran transfection method only suitable for transient transfection?

DEAE Dextran has cytotoxic effects and must be reduced temporarily

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Why does transfection using lipofection work better in rapidly growing and dividing cells?

There is no nuclear envelope to penetrate

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How do you activate dendrimers?

Removal of tertiary amines

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During microinjection, where do you inject the exogenous DNA?

Either into cytoplasm or directly into the nucleus

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Cell Culture - Lecture 8: Gene Transfer at Hochschule Bonn-Rhein-Sieg

Flashcards and summaries for Cell Culture - Lecture 8: Gene Transfer at the Hochschule Bonn-Rhein-Sieg

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