Application of genetic engineeeeeeeringggggggggggg Flashcards
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What do transient transfections allow?
The conduction of short term examinations
What do stable transfections allow?
The conduction of long term examinations
What is used to do a transient transfection?
A liposome
What is a liposome?
A spherical molecule that consists of a lipid bilayer and an aqueous interior
What is the aqueous interior of a lipsome used for?
packaging nucleic acids into
How do the contents of a liposome enter a cell?
The lipsome readily fuses w/ the cell membrane
Why can the lipsome fuse with the cell membrane?
Both the cell and lipsome membranes are lipid bilayers
How are nucleic acids put into a liposome?
They are mixed with the lipofection reagent and left for a short amount of time
Why is transient transfection only short term?
The DNA isn’t bound to histones, so nucleases will degrade the foreign DNA
Why is stable transfection long term?
The DNA beng transfected is integrated into the nucleus
Why does stable transfection take longer than transient transfection
The use of retrovirus-mediated infection takes weeks to performm
What makes up a a retrovirus infector?
RNA genome, reverse transcriptase and integrase all packaged into a viral particle (kinda like HIV)
How do retroviral particles work?
Binds to receptor on target cell, delivers the RNA genome and two enzymes. RT makes DNA copy of RNA, and integrase puts that DNA into the host genome
How are retroviral particles made?
A packaging cell is used
How does a packaging cell make a retroviral particle?
Vectors are transfected into the packaging cell which expresses all the components required for the retroviral particle except the RNA genome
