Spatially Targeted Transfection of Mammalian Cells
Mammalian cell transfection with DNA, RNA, small molecules, and proteins is a widely used molecular biology technique. The primary method of cell transfection is electropermeabilization, which increases the permeability of the cell membrane, allowing chemicals, drugs or DNA to be introduced into the cell. Bulk electroporation has been used for DNA transfection in suspension cells but requires strong electric fields, which leads to relatively low-viability rates.
We have developed localized electroporation and gene transfection of mammalian cells using a Janus particle (JP) bead that is magnetically and electrically powered, leading to the localized, single-cell, successful transfection with high viability rates.
ADVANTAGES OVER EXISTING TECHNOLOGIES
• Maneuvering JP particle to the desired location within closed chambers.
• Method of label-free cargo (e.g. plasmids) loading by dielectrophoresis.
• Enables the use of lower electrical fields for successful electroporation and hence resulting in increased cell viability.
• The use of Janus particle (JP) enables the on/off loading of cargo (e.g. plasmids, cells, organelles etc.) upon request by controlling the electrical field.
• Single-cell localized spatial electroporation of mammalian cells (i.e., targeting the dendrites or axon in a neuron cell).
• AI may be used to automatically manipulate the JP particles to offload their cargo at desired locations, target specific cells and control their interaction with it.
• High-throughput applications using many such JPs and many cells is possible.
US and EU patents pending
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Wu, Y., Fu, A., & Yossifon, G., Active particles as mobile microelectrodes for selective bacteria electroporation and transport. Science Advances, 6(5), 1–12 (2020).
Wu, Y., Fu, A., & Yossifon, G., Micromotor-based localized electroporation and gene transfection of mammalian cells, Proc. Natl. Acad. Sci. 118:38 e2106353118 (2021).