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RET II Presentation. DOTAP Cationic Lipid. DOPC Neutral Lipid. DOPE Neutral Lipid. Lipid Stock TAP/PC & TAP/PE. RET I Research: Gene Therapy. Self-Assembly. Driving Force: HYDROPHOBIC EFFECT. Mixture of Cationic And Neutral Lipids. sss. ss. ss. s. sss. SSss. ss. sss. ss.
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DOTAP Cationic Lipid DOPC Neutral Lipid DOPE Neutral Lipid Lipid StockTAP/PC & TAP/PE RET I Research: Gene Therapy
Self-Assembly Driving Force: HYDROPHOBIC EFFECT Mixture of Cationic And Neutral Lipids sss ss ss s sss SSss ss sss ss ss sssS Ss ss sss ss sss sss ss ss Complex Formation Lipids:Liposomes: Polar Head group Glycerol Backbone Hydrocarbon Chains
DNA Liposome Complex sss ss ss sss s ss SSss sss ss ss Ss sssS ss sss ss sss sss ss ss Forming Complexes of DNA and Lipids Complex Formation:
Transfecting Cells Cells: Mouse Fibroblast L-cells Complex Formation
Transfection Results 10 Fold Increase in Transfection Efficiency
2a. 1. 2b. 3. 4. Possible Pathways for releasing DNA • PE is able to release DNA from endosome (4), while PC is trapped in endosome (3) B. PE fuses directly with cell membrane
RET II: Lesson Plans Genetic Engineeringactivities 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
Thank Yous! • Martina • Melanie • Fellow RETers • NIH • Nate & Chris