RET II Presentation
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Transcript of RET II Presentation
RET II Presentation
Lipid Stock TAP/PC & TAP/PE
DOTAPCationic Lipid
DOPCNeutral Lipid
DOPENeutral Lipid
RET I Research: Gene Therapy
Driving Force: HYDROPHOBIC EFFECT
Self-Assembly
HydrocarbonChains
Glycerol Backbone
Polar Head group
Mixture of CationicAnd Neutral Lipids
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Lipids: Liposomes:
Complex Formation
Forming Complexes of DNA and Lipids
Complex Formation:
DNA
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Liposome Complex
Transfecting Cells
Complex Formation Cells: Mouse Fibroblast
L-cells
Transfection Results
DOTAP in DOPC or DOPE - Sub-sectioned
1.00E+03
1.00E+04
1.00E+05
1.00E+06
10 15 20 25 30 35 40 45
Mole % DOTAP
TR
AN
SF
EC
TIO
N E
FF
ICIE
NC
Y
Re
lati
ve
Lig
ht
Un
its
(R
LU
)/m
g p
rote
in
DOTAP/DOPC
DOTAP/DOPE
DOTAP in DOPC or DOPE - Transfection Results
1.00E+03
1.00E+04
1.00E+05
1.00E+06
0 20 40 60 80 100
Mole % DOTAP
TR
AN
SF
EC
TIO
N E
FF
ICIE
NC
Y R
ela
tiv
e
Lig
ht
Un
its
(R
LU
)/m
g p
rote
in
TAP/PC
TAP/PE
10 Fold Increase in TransfectionEfficiency
Possible Pathways for releasing
DNAA. PE is able to
release DNA from endosome (4), while PC is trapped in endosome (3)
B. PE fuses directly with cell membrane
1.
2b.
3.
4.
2a.
RET II: Lesson Plans
Genetic Engineering activities and labs that can be used as a
supplement with current lessons to enhance learning of state standards.
Biology State Standards
– 5a. Students know the general structures and functions of DNA, RNA, and protein.
– 5c. Students know how genetic engineering (biotechnology) is used to produce novel biomedical and agricultural products.
– 5d. Students know how basic DNA technology (restriction digestion by endonucleases, gel electrophoresis, ligation, and transformation) is used to construct recombinant DNA molecules.
Activities
• Genetic Disorder Poster
• Genetically Modified Food Debate
• Genetic Counseling Skit
Labs
• DNA Extractions
• DNA Fingerprinting (Gel Electrophoresis)
• Plant Tissue Differentiation
Simulated Labs
Thank Yous!
• Martina
• Melanie
• Fellow RETers
• NIH
• Nate & Chris