Engineering B-Cells for Antibody Production: Exploring Germinal Center Formation Capacity and Transfection Potential Via Nanowires
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Colon, Aolani
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Abstract
Infectious diseases such as respiratory syncytial virus (RSV), influenza, Epstein-Barr virus (EBV), and human immunodeficiency virus (HIV) have historically been difficult to develop effective vaccines for. However, CRISPR-Cas9 technology offers an alternative approach to induce long-term protection by editing B-cells to produce neutralizing antibodies targeted against these viruses. Building upon a previously established CRISPR model by Moffett et al. (2019), this study aimed to enhance the process involved in engineering B-cells for RSV. To explore potential alternatives to nucleofection, we investigated the use of nanowire technology, previously successful in T-cell transfection, for B-cell engineering. A comprehensive comparison between nanowire-mediated and nucleofection transfection methods was conducted to assess their impact on engineered B-cells (EBCs). Remarkably, nanowire-mediated transfection achieved editing efficiency on par with nucleofection, while mitigating cell death and eliminating the need for pre-activation. Notably, RSV-EBCs transfected using nanowires displayed enhanced germinal center markers, indicative of advanced B-cell maturation processes. This study represents the first successful application of nanowire technology in B-cells and demonstrates its capability of achieving efficient CRISPR knock-ins.
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2023-07-31
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