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

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Study with flashcards and summaries for the course Cell Culture - Lecture 8: Gene Transfer at the Hochschule Bonn-Rhein-Sieg

Exemplary flashcards for Cell Culture - Lecture 8: Gene Transfer at the Hochschule Bonn-Rhein-Sieg on StudySmarter:

What are the two types of transfection?

Exemplary flashcards for Cell Culture - Lecture 8: Gene Transfer at the Hochschule Bonn-Rhein-Sieg on StudySmarter:

For which type of transfection are plasmids used?

Exemplary flashcards for Cell Culture - Lecture 8: Gene Transfer at the Hochschule Bonn-Rhein-Sieg on StudySmarter:

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

Exemplary flashcards for Cell Culture - Lecture 8: Gene Transfer at the Hochschule Bonn-Rhein-Sieg on StudySmarter:

By which mechanism is exogenous DNA internalized into target cells?

Exemplary flashcards for Cell Culture - Lecture 8: Gene Transfer at the Hochschule Bonn-Rhein-Sieg on StudySmarter:

Name three chemical procedures to introduce DNA into cells.

Exemplary flashcards for Cell Culture - Lecture 8: Gene Transfer at the Hochschule Bonn-Rhein-Sieg on StudySmarter:

For what reason are transfection reagents positively charged?

Exemplary flashcards for Cell Culture - Lecture 8: Gene Transfer at the Hochschule Bonn-Rhein-Sieg on StudySmarter:

What are three disadvantages about using Calcium Phosphate for transfection?

Exemplary flashcards for Cell Culture - Lecture 8: Gene Transfer at the Hochschule Bonn-Rhein-Sieg on StudySmarter:

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.

Exemplary flashcards for Cell Culture - Lecture 8: Gene Transfer at the Hochschule Bonn-Rhein-Sieg on StudySmarter:

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

Exemplary flashcards for Cell Culture - Lecture 8: Gene Transfer at the Hochschule Bonn-Rhein-Sieg on StudySmarter:

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

Exemplary flashcards for Cell Culture - Lecture 8: Gene Transfer at the Hochschule Bonn-Rhein-Sieg on StudySmarter:

How do you activate dendrimers?

Exemplary flashcards for Cell Culture - Lecture 8: Gene Transfer at the Hochschule Bonn-Rhein-Sieg on StudySmarter:

During microinjection, where do you inject the exogenous DNA?

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Exemplary flashcards for Cell Culture - Lecture 8: Gene Transfer at the Hochschule Bonn-Rhein-Sieg on StudySmarter:

Cell Culture - Lecture 8: Gene Transfer

What are the two types of transfection?

  • Transient transfection
  • Stable transfection

Cell Culture - Lecture 8: Gene Transfer

For which type of transfection are plasmids used?

Transient

Cell Culture - Lecture 8: Gene Transfer

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

  • Lysosome/Endosome
  • Nucleases

Cell Culture - Lecture 8: Gene Transfer

By which mechanism is exogenous DNA internalized into target cells?

Endocytosis

Cell Culture - Lecture 8: Gene Transfer

Name three chemical procedures to introduce DNA into cells.

  • Calcium Phosphate
  • DEAE Dextran
  • Artificial liposomes

Cell Culture - Lecture 8: Gene Transfer

For what reason are transfection reagents positively charged?

To bind to negatively charged DNA backbone

Cell Culture - Lecture 8: Gene Transfer

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)

Cell Culture - Lecture 8: Gene Transfer

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.

Cell Culture - Lecture 8: Gene Transfer

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

DEAE Dextran has cytotoxic effects and must be reduced temporarily

Cell Culture - Lecture 8: Gene Transfer

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

There is no nuclear envelope to penetrate

Cell Culture - Lecture 8: Gene Transfer

How do you activate dendrimers?

Removal of tertiary amines

Cell Culture - Lecture 8: Gene Transfer

During microinjection, where do you inject the exogenous DNA?

Either into cytoplasm or directly into the nucleus

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