title: Comparison between direct and reverse electroporation of cells in situ: a simulation study.
abstract: The discovery of the human genome has unveiled new fields of genomics, transcriptomics, and proteomics, which has produced paradigm shifts on how to study disease mechanisms, wherein a current central focus is the understanding of how gene signatures and gene networks interact within cells. These gene function studies require manipulating genes either through activation or inhibition, which can be achieved by temporarily permeabilizing the cell membrane through transfection to delivercDNAorRNAi. An efficient transfection technique is electroporation, which applies an optimized electric pulse to permeabilize the cells of interest. When the molecules are applied on top of seeded cells, it is called "direct" transfection and when the nucleic acids are printed on the substrate and the cells are seeded on top of them, it is termed "reverse" transfection. Direct transfection has been successfully applied in previous studies, whereas reverse transfection has recently gained more attention in the context of high-throughput experiments. Despite the emerging importance, studies comparing the efficiency of the two methods are lacking. In this study, a model for electroporation of cells in situ is developed to address this deficiency. The results indicate that reverse transfection is less efficient than direct transfection. However, the model also predicts that by increasing the concentration of deliverable molecules by a factor of 2 or increasing the applied voltage by 20%, reverse transfection can be approximately as efficient as direct transfection.
authors.affiliation: Department of Bioengineering, Imperial College London, London, United Kingdom.
authors.forename: Leila
authors.initials: L
authors.lastname: Towhidi
authors.affiliation: Department of Bioengineering, Imperial College London, London, United Kingdom.
authors.forename: Delaram
authors.initials: D
authors.lastname: Khodadadi
authors.affiliation: Department of Bioengineering, Imperial College London, London, United Kingdom.
authors.forename: Nataly
authors.initials: N
authors.lastname: Maimari
authors.affiliation: Department of Bioengineering, Imperial College London, London, United Kingdom.
authors.forename: Ryan M
authors.initials: RM
authors.lastname: Pedrigi
authors.affiliation: Department of Bioengineering, Imperial College London, London, United Kingdom.
authors.forename: Henry
authors.initials: H
authors.lastname: Ip
authors.affiliation: Department of Bioengineering, Imperial College London, London, United Kingdom.
authors.forename: Zoltan
authors.initials: Z
authors.lastname: Kis
authors.affiliation: Department of Pathology and Immunology, University of Geneva, Geneva, Switzerland Department of Medical Specializations - Cardiology, University of Geneva, Geneva, Switzerland.
authors.forename: Brenda R
authors.initials: BR
authors.lastname: Kwak
authors.affiliation: Department of Oncology, Lausanne University Hospital and University of Lausanne, Lausanne, Switzerland.
authors.forename: Tatiana W
authors.initials: TW
authors.lastname: Petrova
authors.affiliation: Department of Oncology, Lausanne University Hospital and University of Lausanne, Lausanne, Switzerland.
authors.forename: Mauro
authors.initials: M
authors.lastname: Delorenzi
authors.affiliation: Department of Bioengineering, Imperial College London, London, United Kingdom r.krams@imperial.ac.uk.
authors.forename: Rob
authors.initials: R
authors.lastname: Krams
mesh_terms: D000818:Animals; D001692:Biological Transport; D002462:Cell Membrane / Q000378:metabolism*; D002463:Cell Membrane Permeability; D002478:Cells, Cultured; D003198:Computer Simulation; D004058:Diffusion; D018274:Electroporation; D042783:Endothelial Cells / Q000378:metabolism*; D006801:Humans; D008957:Models, Genetic; D009716:Numerical Analysis, Computer-Assisted; D016062:Porosity; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000378:metabolism*; D012984:Software; D014162:Transfection / Q000379:methods*
chemical_list: D034741:RNA, Small Interfering
keywords: Electroporation; high‐throughput techniques; transfection efficiency