Physical methods of transfection other than electroporation include the biolistic particle delivery system, direct microinjection, and laser-mediated gene transfer. Although these physical methods differ in the tools they employ, they all enable the direct transfer of nucleic acids into the cytoplasm or the nucleus by membrane penetration without using chemicals or viruses.
Biolistic particle delivery system
In brief, the biolistic particle delivery system, also known as particle bombardment, involves projecting microscopic heavy-metal particles (often gold or tungsten) coated with nucleic acids into recipient cells at high velocity using a ballistic device (i.e., “gene gun”). Biolistic particle delivery can be used to transiently transfect dividing and non‑dividing cells in culture as well as cells in vivo, and it is often used for genetic vaccination and agriculture applications [1–3].
While this technique is reliable and fast, it requires costly equipment, causes physical damage to the samples, and necessitates high cell numbers due to high mortality rates.
Microinjection method of gene transfer
The microinjection method of gene transfer delivers nucleic acids into the cytoplasm or the nucleus one cell at a time by means of a fine needle. Therefore, this method is limited to ex vivo applications such as the transfer of genes into oocytes to engineer transgenic animals or the delivery of artificial chromosomes [4–6].
Although direct microinjection is nearly 100% efficient, it demands considerable technical skill, is extremely labor-intensive, and often causes cell death. As such, this method is not appropriate for studies that require the transfection of large numbers of cells.
Laser-mediated gene transfer
Laser-mediated gene transfer, also known as phototransfection, laserfection, or optoporation, uses a laser pulse to transiently permeabilize the cell membrane [7–8]. When the laser induces a pore in the membrane, the osmotic difference between the medium and the cytosol facilitates the entry of nucleic acids or other desired substances in the medium (ions, small molecules, proteins, semiconductor nanocrystals, etc.) into the cell.
Advantages of laser-mediated transfection include high transfection efficiency and the ability to make pores at any location on the cell. However, this method requires an expensive laser-microscope system and the cells to be attached to a substrate.
In addition to the physical methods of transfection mentioned above, other physical delivery technologies use hydrodynamic pressure, ultrasound, or magnetic field to drive naked nucleic acids or nucleic acid-particle complexes into recipient cells.
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- Klein TM, Arentzen R, Lewis PA, Fitzpatrick-McElligott S. Transformation of microbes, plants and animals by particle bombardment. Biotechnology (N Y). 1992 Mar;10(3):286–291. doi: 10.1038/nbt0392-286. PMID: 1368100.
- Ye GN, Daniell H, Sanford JC. Optimization of delivery of foreign DNA into higher-plant chloroplasts. Plant Mol Biol. 1990 Dec;15(6):809–819. doi: 10.1007/BF00039421. PMID: 2103474.
- Burkholder JK, Decker J, Yang NS. Rapid transgene expression in lymphocyte and macrophage primary cultures after particle bombardment-mediated gene transfer. J Immunol Methods. 1993 Oct 15;165(2):149–156. doi: 10.1016/0022-1759(93)90340-d. PMID: 8228267.
- Capecchi MR. High efficiency transformation by direct microinjection of DNA into cultured mammalian cells. Cell. 1980 Nov;22(2 Pt 2):479–488. doi: 10.1016/0092-8674(80)90358-x. PMID: 6256082.
- Capecchi MR. Altering the genome by homologous recombination. Science. 1989 Jun 16;244(4910):1288–1292. doi: 10.1126/science.2660260. PMID: 2660260.
- Telenius H, Szeles A, Keresö J, Csonka E, Praznovszky T, Imreh S, Maxwell A, Perez CF, Drayer JI, Hadlaczky G. Stability of a functional murine satellite DNA-based artificial chromosome across mammalian species. Chromosome Res. 1999;7(1):3-7. doi: 10.1023/a:1009215026001. PMID: 10219727.
- Shirahata Y, Ohkohchi N, Itagak H, Satomi S. New technique for gene transfection using laser irradiation. J Investig Med. 2001 Mar;49(2):184–190. doi: 10.2310/6650.2001.34045. PMID: 11288759.
- Schneckenburger H, Hendinger A, Sailer R, Strauss WS, Schmitt M. Laser-assisted optoporation of single cells. J Biomed Opt. 2002 Jul;7(3):410–416. doi: 10.1117/1.1485758. PMID: 12175291.
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