RET II Presentation
RET I Research: Gene Therapy
Lipid Stock
TAP/PC & TAP/PE
DOTAP
Cationic Lipid
DOPC
Neutral Lipid
DOPE
Neutral Lipid
Complex Formation
Lipids:
Liposomes:
Self-Assembly
sss
Polar
Head group
Glycerol
Backbone
Hydrocarbon
Chains
Driving Force:
HYDROPHOBIC EFFECT
Mixture of Cationic
And Neutral Lipids
Forming Complexes of
DNA and Lipids
Complex Formation:
sss
DNA
Liposome
Complex
Transfecting Cells
Complex Formation
Cells:
Mouse Fibroblast
L-cells
Transfection Results
DOTAP in DOPC or DOPE - Transfection Results
TAP/PC
TAP/PE
DOTAP/DOPC
DOTAP/DOPE
DOTAP in DOPC or DOPE - Sub-sectioned
1.00E+06
1.00E+05
1.00E+04
1.00E+03
0
20
40
60
Mole % DOTAP
80
100
TRANSFECTION EFFICIENCY
Relative Light Units (RLU)/mg protein
TRANSFECTION EFFICIENCY Relative
Light Units (RLU)/mg protein
1.00E+06
1.00E+05
1.00E+04
10 Fold Increase in Transfection
Efficiency
1.00E+03
10
15
20
25
30
Mole % DOTAP
35
40
45
Possible Pathways
for releasing
DNA
A. PE is able to
release DNA
from endosome
(4), while PC is
trapped in
endosome (3)
B. PE fuses
directly with
cell membrane
2a.
1.
2b.
3.
4.
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