The document outlines various methods for transferring DNA into eukaryotic host cells, including chemical transfection, electroporation, microinjection, and gene gun techniques. Each method has its specific applications and advantages, such as the efficiency of liposome-mediated transport and the direct targeting of cells with microinjection. However, challenges like efficiency dependency on cell health and high costs of certain methods are also noted.

1. METHODS TO TRANSFER
DNA TO HOST CELL

2. METHODS
Different methods of introducing genes to the eukaryotic host
cell
• Chemical
transfection
• Lipofection
Chemical
method
• Electroporation
• Microinjection
• Gene gun
Physical
method

3. Transfection of Eukaryotic Cells

4. Transfection
 Transfection is a method of transporting DNA, RNA
and/or various macromolecules into an eukaryotic
cell
 Using chemical, lipid or physical based methods
 Neutralize negative charges on DNA

5. TYPES
Chemical method:
 Calcium phosphate creates
precipitates that settle on cells
and are taken in
Lipid or Polymer method:
 Interact with DNA, promotes
binding to cells and uptake via
endocytosis

6. TRANSFECTION

7. Purpose of Transfection
 Study gene function
 Study protein expression
 Transfer DNA into embryonic stem cells

8. Electroporation
 Use of high voltage to deliver nucleic acids
 Pores are formed on cell membrane

9. Electroporation

11. Applications
 Direct transfer of plasmid between cells
 Transdermal drug delivery
 Cancer tumour electro chemotherapy
 Gene therapy

12. Microinjection
 Use of a fine needle
 Use to transfer of DNA into embryonic stem cells
 Injection of linear DNA
 Random integration
 Inefficient

14. Advantages
 No requirement of marker gene
 Introduction of the target gene directly into a single cell
 Easy identification of transformed cells
 No requirement of selection of transformed cells
(antibiotic resistance or herbicide resistance markers)
 Can be used for transgenic animals

15. Gene gun
Biolistic Particle delivery
 Uses high velocity for delivery of nucleic acids & penetration of cell wall
 Also termed as particle bombardment ,particle gun, microprojectile

17. Advantages
 Simple and convenient method involving coating DNA/RNA on to gold
micro-carrier, loading sample cartridges, pointing the nozzle and firing the
device.
 Eliminates the use of potentially harmful viruses or toxic chemical
treatment as gene delivery vehicle.

18. Advantages & Disadvantages
Advantages
 Provides the ability to transfer
in negatively charged
molecules into cells with a
negatively charged membrane
 Liposome-mediated transport
of DNA has high efficiency.
Good for long-term studies
requiring incorporation of
genetic material into the
chromosome
Disadvantages
 Chemical Reagents: not useful
for long-term studies
 Transfection efficiency is
dependent on cell health, DNA
quality, DNA quantity,
confluency (40-80%)
 Direct Microinjection and
Biolistic Particle delivery is an
expensive and at times a
difficult method