Microcarrier particles (ex. gold, tungsten) are coated with recombinant DNA. A pressurized helium pulse or electric discharge is used to deliver the DNA-coated particles at a high velocity through the cell membrane of target cells. The gene gun can also be used to penetrate plant cell walls.

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Non-viral vectors deliver genetic material into cells and have transient effects and lower efficiency than viral vectors, but offer many advantages: low immunogenicity (the ability to provoke an undesirable immune response), larger DNA capacity, and lower cost. Examples of non-viral vectors include: naked DNA, physical methods (electroporation, sonoporation, gene gun, magnetofection, hydroporation), and chemical carriers.

Non-viral vectors

- Electroporation
- Microinjection
- Gene Guns
- Shuttle Vectors
- Viral Vectors

Techniques for Transfection in Eukaryotic Cells

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Naked plasmid DNA is injected into the body and enters cells through endocytosis. This method is simple and safe but lacks efficiency.

Naked DNA

An electric field greater than membrane capacitance is applied, causing the membrane to break down and form a pore, through which genetic material can pass. The pores can be temporary or permanent, depending on the strength and duration of the stimulus.

Electroporation

Ultrasound waves are applied to make the cell membrane permeable to DNA , allowing genetic material to pass into the cell. This mechanism involves microbubbles, which oscillate in response to ultrasound waves and create an opening in the membrane.

Sonoporation

Water pressure is applied to the cell membrane to create openings in the membrane, allowing genetic material to pass into the cell. The pressure is created by injecting a high volume of DNA solution.

Hydroporation

The process of transferring genes bound by magnetic nanoparticles (ex. iron oxide) into cells. An external magnetic field is applied below the cells, and genetic material is transferred into cells by endocytosis. An advantage of magnetofection is that the cell membrane is not physically damaged to form pores. 

Magnetofection

Gene gun

Eukaryotic cells can be transfected by electric pulses that disrupt the cell membrane to create pores where genes can be introduced. A positive charge is also applied inside the cell to pull in DNA.

Eukaryotic Electroporation

Recombinant DNA can be injected into the eukaryotic cells directly by using a microinjection needle.

Microinjection

Shuttle vectors are plasmids that can be transfected between bacteria and eukaryotic cells. 

Shuttle Vectors 

Viral vectors use inactive versions of viruses to deliver genetic material into cells, taking advantage of viruses' ability to easily enter cells. They are the most common vector type in gene therapy because they produce long-term, high level of gene expression. Examples include: adeno-associated viral vectors (AAVs), adenoviral vectors (AdVs), and lentiviral vectors (LVVs).

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