Meta-Analysis of Transfection

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Transfection is a critical technique for introducing nucleic acids—such as DNA, RNA, or oligonucleotides—into cells and plays a pivotal role in diverse research fields, including gene therapy, recombinant protein production, and functional genomics. This meta-analysis examines the fundamental mechanisms, methodologies, and challenges associated with transfection, highlighting advances in both viral and non-viral delivery systems, optimization techniques, and clinical applications. Viral vectors, including adenovirus, retrovirus, and lentivirus, offer high efficiency and are frequently used in gene therapy applications, while non-viral methods, such as lipid-mediated transfection, polyethylenimine (PEI), dendrimer complexes, electroporation, microinjection, and biolistic delivery, provide safer alternatives but often exhibit lower efficiency, necessitating optimization. Lipid-based transfection remains one of the most widely used methods, particularly lipofection, due to its ease of use and efficiency in commonly used cell lines like HEK293 and HeLa cells. Electroporation is effective for challenging cell types, such as primary neurons and stem cells, though high cell mortality rates necessitate careful optimization. Transfection has significantly contributed to gene therapy, particularly for genetic disorders such as cystic fibrosis, hemophilia, and spinal muscular atrophy, with viral vectors employed in CAR-T cell therapy for cancer treatment showing promising results in hematological malignancies. Recent advances in mRNA transfection have revolutionized vaccine development, exemplified by mRNA vaccines for COVID-19, demonstrating the potential for further therapeutic applications. However, challenges remain, including achieving high transfection efficiency while maintaining cell viability, especially in primary cells and stem cells, which are more resistant to transfection than immortalized cell lines. Cytotoxicity and off-target effects limit the clinical utility of transfection, particularly in gene therapy, where insertional mutagenesis poses significant safety risks. The cost of scaling transfection for therapeutic applications, such as CAR-T cell production, remains prohibitive. Future research will focus on overcoming these limitations while advancing the clinical applications of transfection for therapeutic gene delivery and personalized medicine. Overall, as new materials, technologies, and optimization strategies are developed, the efficacy, safety, and applicability of transfection techniques will likely improve, enhancing their role as cornerstones of molecular biology and biotechnology.