An efficient non-viral gene delivery for varieties of cells has been considered essential for gene therapy and tissue engineering. This study evaluated transfection efficiency of chitosan (HW) with molecular weights (Mw) at 470 and degree of deacetylation (DDA) 80% and its depolymerization product (LW) with Mw at 16 kDa and DDA 54%, as well as epidermal growth factor (EGF) conjugated to chitosan-DNA microparticles of both HW and LW by using either disulfide linkage or NHS-PEO(4)-Maleimide as a cross linker. The results revealed that the depolymerized LW at chitosan/DNA charge ratio 56:1 and pH 6.9 gave high transfection efficiency in both KB, a cancer cell line, and fibroblast cells at about the same level of Lipofectamine, but the EGF-conjugated chitosan-DNA polyplexes from these methods did not improve transfection efficiency, which may come from the aggregation and fusing of the complexes as shown in scanning electron microscopy. However, this depolymerized LW chitosan showed the potential for further development as a safe and cost-effective non-viral gene delivery vehicle.
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http://dx.doi.org/10.1007/s10856-010-3993-9 | DOI Listing |
Lipid nanoparticles (LNP) are the most clinically advanced non-viral gene delivery system. While progress has been made for enhancing delivery, cell specific targeting remains a challenge. Targeting moieties such as antibodies can be chemically-conjugated to LNPs however, this approach is complex and has challenges for scaling up.
View Article and Find Full Text PDFBr J Haematol
January 2025
Institute of Hematology, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, Hubei, China.
Chimeric antigen receptor T-cell (CAR-T) therapy has shown transformative potential in treating malignant tumours, with increasing global approval of CAR-T products. However, high-production costs and risks associated with viral vector-based CAR-T cells-such as insertional mutagenesis and secondary tumour formation-remain challenges. Our study introduces an innovative CAR-T engineering approach using mRNA delivered via lipid nanoparticles (LNPs), aiming to reduce costs and enhance safety while maintaining strong anti-tumour efficacy.
View Article and Find Full Text PDFHum Gene Ther
January 2025
School of Bioengineering, East China University of Science and Technology, Shanghai, China.
Adeno-associated virus (AAV)-associated gene therapy has been increasingly promising, in light of the drugs progressed to clinical trials or approved for medications internationally. Therefore, scalable and efficient production of recombinant AAV is pivotal for advancing gene therapy. Traditional methods, such as the triple-plasmid transfection of human embryonic kidney 293 cells in suspension culture, have been widely employed but often hampered by low unit yield.
View Article and Find Full Text PDFMessenger RNA (mRNA) therapeutics have garnered considerable attention due to their remarkable efficacy in the treatment of various diseases. The COVID-19 mRNA vaccine and RSV mRNA vaccine have been approved on the market. Due to the inherent nuclease-instability and negative charge of mRNA, delivery systems are developed to protect the mRNA from degradation and facilitate its crossing cell membrane to express functional proteins or peptides in the cytoplasm.
View Article and Find Full Text PDFJ Virol Methods
January 2025
Huzhou Key Laboratory of Innovation and Application of Agricultural Germplasm Resources, Huzhou Academy of Agricultural Sciences, Huzhou 313000, China. Electronic address:
In this study, a novel Encephalomyocarditis virus (EMCV) reverse genetic operating system was developed utilizing CMV promoters, enabling EMCV genome expression under the transcriptional control of the CMV immediate early promoter and BGH polyA transcriptional-termination signal. The full-length cDNA of EMCV BJC3 was ligated to the pRK5 vector, incorporating the CMV eukaryotic promoter sequence, resulting in the construction of recombinant plasmid EMCV (pEMCV). Subsequently, the recombinant plasmid was transfected into BHK-21 cells to generate the rescue virus.
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