Magnetofection

Question 1

What, por que, is magnetofection?

Answer 1

Magnetically targeted gene transfection

Question 2

What, por que que is gene transfection?

Answer 2

Process of introducing biomaterials into the cell eg DNA/RNA/therapeutic agents/diagnostic agents

Question 3

What are the 4 ways transfection can be accomplished?

Answer 3

1)biochemical methods 2)physical methods 3)virus mediated methods 4)a combo of the above

Question 4

What is gene therapy?

Answer 4

A method to insert genetic material into an individuals cells and tissues in order to replace a defective gene, or by introducing a new function into cells to attempt to treat more complex inherited or aquired genetic disorders such as cancer, immunodeficiency syndrome, cys fib and haemophilia

Question 5

Which cells do we want to target?

Answer 5

Both healthy and cancerous

Question 6

What do we want to target in cancerous cells?

Answer 6

Apoptosis genes

Question 7

What is the ex vivo strategy for transgene delivery

Answer 7

Genetic alterations of patients target cells happenind outside of the body in a culture, target cells from the patient are transfected with the desired therapeutic culture. These modified genes are then reintroduced into the patients body, where they produce the needed proteins that correspond in the inserted gene.

Question 8

What is the in vivo strategy for transgene delivery?

Answer 8

Introduction of genetic material directly into the patients body in the form of a vector (viral, bac plasmid, nans) by cell-specific direct injection into the tissue in need. Once inside and in contact with specifically targeted cells, genetic material incorporate where it encodes produciton of needed protein.

Question 9

What are 2 obstacles to the delivery of material into cells?

Answer 9

1)cells crowded with organelles 2) membrane barrier

Question 10

What are 6 goals for delivering material into cells?

Answer 10

1)dont kill cell 2)dont unrecoverably damage 3) dont adversely change 4) controllable delivery of material 5) vector cant b toxic 6) safe/efficient

Question 11

What is the viral transfection method?

Answer 11

Using viruses to modify cells by delivering DNA

Question 12

What are the two forms of non-viral transfection?

Answer 12

Physical and chemical

Question 13

What are four physical methods?

Answer 13

1)electroporation 2)laser/beam mediated 3)direct injection of naked DNA 4)gene gun

Question 14

What are 4 chemical methods?

Answer 14

1)liposome 2)polymers 3)cationic lipids 4)calcium phosphate

Question 15

What are advantages to non-viral transfection?

Answer 15

Eliminates safety concernes, a variety of techniques and can be used for large groups of cells OR individual cells

Question 16

What are disadvantages to non-viral transfection?

Answer 16

less efficient than viral, technically demanding and no best method for all application

Question 17

How can we tell if the transfection was successful if the changes are not immediately dectectable?

Answer 17

a ‘reporter gene’ tandem construct eg modify a cell so that it internally produces and expresses a green flourescent protein

Question 18

What is the luciferase assay?

Answer 18

a fucking badass name thats what

Question 19

No srsly what is the luciferase assay?

Answer 19

Luciferase is an enxyme that catalyzes the reaction of luciferin with oxygen to produce light. Transfect cells w/luciferase gene and perform assay: extract luciferase via cell lysis, buffer and add components to increase sensitivity and maintain pH, luminescent reaction triggered by an injection of luciferin - emitted light recorded

Question 20

What does magnetofection rely on?

Answer 20

selection of appropriate size and surface coating of magnetic nanoparticles for binding with the gnetic materials. genetic materials may be attached to surface of nanoparticles by employing cleavable linkers/utilizing electrostatic interactions.