Genomics and Applied Biology 2024, Vol.15, No.2, 89-98 http://bioscipublisher.com/index.php/gab 93 Figure 2 Nanocapsules developed for the transfer of genetic material to primary T cells and hematopoietic stem cells (Adopted from Tarakanchikova et al., 2019) Image caption: A) Scanning electron microscopy of the CaCO3 core (left panel), and ready-to-use capsules (middle panel). Scale bar 100 nm. Nanoparticle tracking analysis shows the size distribution between 50 and 280 nm with a peak of 160 nm (right panel). B) Flow cytometry analysis of T cells (upper panels) and CD34+ (bottom panels) cells treated with Rhodamine-labeled nanocapsules showed that application of 5 nanocapsules/cells is sufficient for an efficient transfer of capsules into CD34+ cells, and 10 nanocapsules/cell is sufficient for an efficient transfer of capsules into T cells . Application of higher nanocapsules number led to the reduction of cell viability (left panel). C) Confocal microscopy of T cells and CD34+ cells revealed efficient transfer of GFP mRNA. Images were taken 72 h post-treatment with capsules. Cells were stained with DAPI to visualized cell nuclei. Green fluorescence show cells expressing GFP after uptake of nanocapsules. Scale bar left panel 30 µm, right panel 9 µm (Adopted from Tarakanchikova et al., 2019) 5.2 Case study: development of non-viral gene transfer techniques Non-viral gene transfer techniques have also seen significant progress. For instance, the use of DNA nuclear targeting sequences, such as 3NFs, has been shown to enhance the nuclear import of plasmid DNA, thereby improving gene transfer efficiency in both in vitro and in vivo settings (Guen et al., 2021). This method leverages the interaction with the transcription factor NF-κB to facilitate the translocation of nucleic acids into the nuclear compartment of target cells. Another notable development is the application of carbon dot-polyethyleneimine
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