Comparison of various transfection methods for gene transfection-Gene Transfection
Transfection methods | principle | Main applications | characteristic | Main manufacturers and products | ||
DEAE glucan method | The complex formed by the interaction between positively charged DEAE glucan and negatively charged phosphate backbone of nucleic acid is internalized by cells. | transient transfection | Relatively simple and reproducible compared to calcium phosphate, but it has certain toxic side effects on cells. Serum needs to be removed during transfection and is generally only used for BSC-1, CV-1, COS cell lines | Sigma-Aldrich (DEAE-Dextran Transfection Kit) | ||
calcium phosphate method | Calcium phosphate DNA complex adsorbed onto cell membrane and internalized by cells | Stable transfection, transient transfection | Not suitable for primary cells (requiring high DNA concentration), easy to operate but with poor reproducibility. Some cells are not suitable. CSCL gradient centrifugation is recommended for cells, as transfection requires a high copy number. | |||
cationic polymer | Positively charged liposomes form complexes with negatively charged phosphate groups in nucleic acids, and then the remaining nuclei on the liposomes bind to the negatively charged nuclei of sialic acid residues on the cell membrane; Another explanation is that it is entered into cells through endocytosis. If the DNA concentration is too high, it neutralizes the surface nuclei of liposomes and reduces their binding ability to cells | Stable transfection, transient transfection, all cells | Easy to use, capable of carrying large fragments of DNA, applicable to various types of naked DNA or RNA, capable of transfecting various types of cells, without immunogenicity. Although highly efficient in vitro gene transfection, in vivo it can be cleared by serum and accumulate in lung tissue, inducing strong anti-inflammatory reactions and leading to high levels of toxicity, which greatly limits its application | Invitrogen （ Lipofectamine 2000 ， Lipofectamine ， Lipofectin ， Lipofectamine Plus ， Cellfectin ） Roche （ Dosper ， DOTAP ， FuGENE 6 ） CPG Biotech Co （ GeneLimo Plus ， GeneLimo Super ） Promega （ Transfast ， Tfx ， Transfectam ） | ||
cationic polymer | Positively charged polymers form positively charged complexes with negatively charged phosphate groups in nucleic acids, which then interact with negatively charged proteoglycans on the cell surface and enter the cell through endocytosis. | Stable transfection, transient transfection, all cells | In addition to the high transfection efficiency, simple operation, wide applicability, and good repeatability of cationic liposomes, it also has the characteristics of high transfection efficiency and low cytotoxicity in vivo, making it a new generation of transfection reagents. | |||
Virus mediated method | Retrovirus (RNA) | It enters the host cell through the interaction between the virus membrane glycoprotein and the receptor on the host cell surface, and then reverses into an enzyme to initiate the synthesis of DNA, which is randomly integrated into the host genome | Stable transfection, specific host cells | Can be used for difficult to transfect cells, primary cells, in vivo cells, etc., but the gene carrying capacity should not be too large (<8kb), Cells need to undergo fission phase, and safety factors need to be considered | ||
Adenovirus (double stranded DNA) | First, it binds to the receptors on the cell surface, and then is engulfed by the cell under the mediation of α v integrin. | Instant transfection, specific host cells | Can be used for difficult to transfect cells, safety factors need to be considered. | |||
Biolistic Particle Transfer Method (Gene Gun Particle Bombardment Method) | The DNA was precipitated using microscopic heavy metal particles, and then the coated particles were projected into the cells using a ballistic device, DNA is gradually released and expressed inside the cell | Transient transfection, stable transfection | Can be used for: human epidermal cells, fibroblasts, lymphocyte lines, and primary cells. | ０ | ||
Microinjection method | Directly inject DNA into the target cell nucleus through microscopic manipulation | Stable transfection, transient transfection | The number of transfected cells is limited, and they are mostly used for engineering or transgenic animal embryonic cells. | ０ | ||
Electroporation method | High pulse voltage disrupts the cell membrane potential, and DNA is introduced through small pores formed on the membrane | Stable transfection, transient transfection, all cells | Widely applicable, in addition to plasmids, it can also be transfected into large genomes (>65kb) but has a high cell death rate, The amount of DNA and cells used is large, and the electroporation experimental conditions need to be optimized according to different cell types, with copy numbers ranging from 1 to 20 | ０ | ||
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