A platform for rna-based and genomic cell engineering: simplifying multiplexed delivery to human primary immune and stem cells

Background & Aim Cell therapy has the potential to revolutionize many areas of unmet clinical needs including cancer, degenerative and metabolic diseases, genetic disorders, infection, inflammation and autoimmunity. Both RNA-based and genomic editing technologies (e.g. CRISPR/Cas9) are actively investigated to design cellular therapies with customized functionalities. One of the key advantages of these new technologies is their highly adaptable nature that enables sophisticated cellular engineering, rational design and rapid pursuit of new therapeutic targets. Methods, Results & Conclusion At Portal, we have developed a gentle silicon membrane-based delivery technology that facilitates mechanoporation of cells and can be easily integrated with existing clinical-scale workflows. Our results in primary immune cells using monocytes, T, B and NK cells demonstrate over 80% efficiency and viability of cellular engineering. CRISPR/Cas9 RNPs, mRNAs and siRNAs have been successfully delivered as single cargos or in a multiplexed fashion, demonstrating the potential for either single- or multi-step cell engineering workflows which can address several targets. Furthermore, data in iPSCs and HSCs have also shown significant potential in stem cell engineering and differentiation capabilities. This technology is currently being implemented for research and clinical scale use - including in an integrated Point-of-Care manufacturing system. With continued progress in simplifying cell engineering using both RNA-based and genomic editing technologies, we aim to unlock vast biological potential in the field of cell therapy while simultaneously reducing or even eliminating safety or manufacturing concerns such as inadvertent genome alteration associated with other delivery modalities such as lentiviral vectors.