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

sss sss

ssss

ssssss

sss sss

sss

sss ssss

ssssss ssSSssSSss

sssSsssS

ssssssssssssssssSsSss

sssssss

sss

sssssss

Lipids: Liposomes:

Complex Formation

Forming Complexes of DNA and Lipids

Complex Formation:

DNA

sss sss

ssss

sssssssss sss

sss

sss ssss

ssssssssSSssSSsssssS

sssS

ssssssssssssssssS

sSsssss

ssssss

sss

sssss

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