A method used to apply an electric current across a cell membrane resulting in temporary “pore” formation enabling the uptake of exogenous molecules found in the medium to either the cytoplasm or into the nucleus, thereby transfecting or transforming the cell. Sachin Mehta Reno Nevada Reno, Nevada

2. Theory
• A method used to apply an electric current
across a cell membrane resulting in temporary
“pore” formation enabling the cell membrane
to be semi-permeable—allowing things to
enter the cell for a time being.

3. Theory, contd.
• Electroporation is a process that is used to
introduce foreign genes into a host cell.
• Electric shocks are used as a mechanism for
introducing new DNA into a host cell by creating
new pores in the plasma membrane of the host
cell.
• The new DNA enters the host cell through the
new pores and is incorporated into the genome
of the new cell.

4. Electroporation Cell Process
http://www.youtube.com/watch?v=TPPudsNVcqE

5. Applications
• Transformation of bacteria,
yeasts, etc.
• Can be used in vivo
or
• In vitro
• Chemotherapy
• Gene Therapy
• Diagnostics

6. Example: Inovio’s Electroporation
Technology
Electroporation uses controlled, millisecond
electrical pulses to create temporary pores in the
cell membrane and allow dramatic cellular uptake
of a synthetic DNA vaccine previously injected into
muscle or skin.
• Technology allows to safely and effectively deliver
DNA-based vaccines.
• Numerous human studies demonstrated best in
class immune responses from DNA vaccines.

7. No electroporation
vs. With
electroporation
Green Light
displays the
proteins created
by the DNA
vaccines delivered
using
electroporation

8. Considerations
• Cell Size
• Temperature
• Composition of Electrodes

9. Optimizing Process
• Use 8 pulses of 100 microseconds at
0.2 V.
• Increasing voltage slowly because
cell death can occur quickly.

10. Advantages vs. Disadvantages
Advantages:
• Versatility: Electroporation is effective
with nearly all cell and species types .
• Efficiency: A large majority of cells take in
the target DNA or molecule. In a study on
electro transformation of E. coli, for
example, 80% of the cells received the
foreign DNA .
• Small Scale: The amount of DNA required
is smaller than for other methods
• In vivo: The procedure may be performed
with intact tissue . A paper published
in Developmental Biology showed the
successful transfer of a DNA construct
with a fluorescent reporter gene into
intact mouse brain tissue.
•
• Image of in
vivo electroporation in a
mouse brain. The mouse
brains (telencephalons) in
these images are expressing
reporter genes (EYFP)
introduced in gene
constructs by
electroporation.

11. Disadvantages
• Cell Damage: If the pulses are of the wrong
length or intensity, some pores may become too
large or fail to close after membrane discharge
causing cell damage or rupture
• Nonspecific Transport: The transport of material
into and out of the cell during the time of electro-
permeability is relatively nonspecific. This may
result in an ion imbalance that could later lead to
improper cell function and cell death

12. Electrochemotherapy
• Specialized drug
delivery to affected site
in organism
• First study in 1991
• Bleomycin
• Cisplatin
http://www.jove.com/video/1038/electrochemotherapy-of-tumours