Senior Scientist, Research and Development STEMCELL Technologies Vancouver, British Columbia, Canada
Disclosure(s):
Ulrike Lambertz, PhD: No relevant disclosure to display
Introduction/Rationale: Current intracellular delivery methods face significant limitations, including a restricted range of compatible molecules and cell types, as well as unintended alterations to cell biology. CellPore™ Transfection System addresses these limitations by using mechanoporation technology to efficiently deliver mRNAs, CRISPR-Cas9 ribonucleoproteins (RNPs), siRNAs, proteins and other molecules to the cytosol.
Methods: Recent platform enhancements, including the introduction of new cartridge sizes, have expanded compatibility to a wider range of cell types. These include unactivated and activated primary immune cells (e.g. T, NK, B and myeloid cells), hematopoietic stem and progenitor cells and pluripotent stem cells, which traditionally have been challenging to reliably engineer via conventional methods.
Results: In the context of immunology applications, we demonstrate efficient RNP-mediated knockout of surface MHC-I (74%) or TCRɑβ (94%) in human T cells, and TIGIT (84%), CD16 (66%), NKG2A (81%) or NKp46 (87%) in human NK cells, with minimal impact to cell viability and function. We also demonstrate delivery of reporter mRNA and siRNA to monocytes and monocyte-derived dendritic cells for enabling transient gene expression and RNAi-mediated gene knockdown.
Conclusion: The CellPore™ system offers a gentle and efficient method to engineer a variety of cell types with a wide range of molecules. The simple and versatile workflow makes this a valuable tool for enabling new areas of research.
