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35 Cards in this Set
- Front
- Back
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Two methods of gene delivery
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1) viral
2) non-viral |
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best thing about viral gene delivery
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it has very high efficiency
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best thing about non-viral gene delivery
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it is much safer than viral
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ADENOVIRUS
genetic material? |
DNA
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ADENOVIRUS
maximum size of insert? |
7.5 kB
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ADENOVIRUS
integration? |
none; uses cell's existing machinery to make protein
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ADENOVIRUS
immune response? |
yes
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ADENOVIRUS
do cells need to be dividing? |
no
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AA VIRUS
genetic material? |
DNA
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AA VIRUS
maximum size of insert? |
5 kB
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AA VIRUS
integration? |
yes, integrates into genome at a specific site
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AA VIRUS
immune response? |
low risk
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AA VIRUS
do cells need to be dividing? |
no
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RETROVIRUS
genetic material? |
RNA
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RETROVIRUS
maximum insert size? |
8 kB
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RETROVIRUS
integration? |
insertion into genome is random
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RETROVIRUS
immune response? |
low risk
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RETROVIRUS
do cells need to be dividing? |
yes
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LENTIVIRUS
genetic material? |
RNA
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LENTIVIRUS
maximum size of insert? |
9 kB
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LENTIVIRUS
integration? |
random, but tend to be safer than normal retroviruses
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LENTIVIRUS
immune response? |
low risk
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LENTIVIRUS
do cells need to be dividing? |
no
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Non-viral gene delivery methods
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- Naked DNA (ex. plasmid)
- lipoplexes/polyplexes |
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PLASMID
genetic material? |
DNA
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PLASMID
maximum size of insert? |
there is no maximum size
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PLASMID
integration? |
will not integrate into genome unless it is modified
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PLASMID
immune resonse? |
none
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Ways to improve naked DNA delivery
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- calcium phosphate: condsenses DNA and increases transfection
- electroporation: electricial pulse forms a temporary hole in the cell membrane (also increases cytotoxicity) - microinjection: deliver DNA directly to nucleus |
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LIPOPLEX
what is it? |
- associate plasmid DNA with positively charged lipids/polymers
- DNA is collapsed - prevents DNA degradation - improves cells' interaction with negative cell surface |
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LIPOPLEX
genetic material? |
DNA
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LIPOPLEX
maximum size of insert? |
there is no maximum size
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LIPOLPLEX
integration? |
does not integrate
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LIPOPLEX
immune response? |
low immune risk
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steps to gene delivery
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1) transport to cell
2) endocytosis 3) escape from endosome 4) transport to nucleus 5) expression |
