Magnetofection Flashcards
What, por que, is magnetofection?
Magnetically targeted gene transfection
What, por que que is gene transfection?
Process of introducing biomaterials into the cell eg DNA/RNA/therapeutic agents/diagnostic agents
What are the 4 ways transfection can be accomplished?
1)biochemical methods 2)physical methods 3)virus mediated methods 4)a combo of the above
What is gene therapy?
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
Which cells do we want to target?
Both healthy and cancerous
What do we want to target in cancerous cells?
Apoptosis genes
What is the ex vivo strategy for transgene delivery
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.
What is the in vivo strategy for transgene delivery?
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.
What are 2 obstacles to the delivery of material into cells?
1)cells crowded with organelles 2) membrane barrier
What are 6 goals for delivering material into cells?
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
What is the viral transfection method?
Using viruses to modify cells by delivering DNA
What are the two forms of non-viral transfection?
Physical and chemical
What are four physical methods?
1)electroporation 2)laser/beam mediated 3)direct injection of naked DNA 4)gene gun
What are 4 chemical methods?
1)liposome 2)polymers 3)cationic lipids 4)calcium phosphate
What are advantages to non-viral transfection?
Eliminates safety concernes, a variety of techniques and can be used for large groups of cells OR individual cells
What are disadvantages to non-viral transfection?
less efficient than viral, technically demanding and no best method for all application
How can we tell if the transfection was successful if the changes are not immediately dectectable?
a ‘reporter gene’ tandem construct eg modify a cell so that it internally produces and expresses a green flourescent protein
What is the luciferase assay?
a fucking badass name thats what
No srsly what is the luciferase assay?
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
What does magnetofection rely on?
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.
