An Overview of Methods and Tools for Transfection of Eukaryotic Cells in vitro

Abstract

Transfection is a powerful analytical tool enabling studies of gene products and functions in eukaryotic cells. Successful delivery of genetic material into cells depends on DNA quantity and quality, incubation time and ratio of transfection reagent to DNA, the origin, type and the passage of transfected cells, and the presence or absence of serum in the cell culture. So far a number of transfection methods that use viruses, non-viral particles or physical factors as the nucleic acids carriers have been developed. Among non-viral carriers, the cationic polymers are proposed as the most attractive ones due to the possibility of their chemical structure modification, low toxicity and immunogenicity. In this review the delivery systems as well as physical, biological and chemical methods used for eukaryotic cells transfection are described and discussed.

Magnetite Nanoparticles in Magnetic Hyperthermia and Cancer Therapies: Challenges and Perspectives

“…So far, several methods of delivering genetic material to the patient’s body have been proposed. Among them are distinguished methods based on viral vectors and physical and chemical methods [ 147 , 151 ]. The use of the vectors proposed so far in cancer gene therapy is associated with immunogenicity, limited genetic burden, and cancer risk due to the insertion of a therapeutic load in the vicinity of genes responsible for controlling cell growth [ 152 ].…”

A live-imaging protocol for tracking receptor dynamics in single cells

“… Note: In this study, we are performing gene expression by electroporation. Other gene delivery methods, such as viruses, can be used for both in vitro and in vivo experiments ( Fus-Kujawa et al., 2021 ). …”

Concatenation of Transgenic DNA: Random or Orchestrated?

“…In theory, copy arrangements there would be more random due to NHEJ/MMEJ domination through the cell cycle [ 88 ], although non-random head-to-tail concatemers [ 85 ] and end recombination (our unpublished data) suggest that concatenated ends could be processes by HR in cells. In addition, transgenic cell clones are obtained by lipofection, electroporation, or viral delivery [ 94 ] in the asynchronous cell cultures. It would be interesting to compare concatemers in embryos/cultured cells obtained by the same delivery method (microinjection).…”