Intracellular transport of electrotransferred DNA cargo is governed by coexisting ergodic and non ergodic anomalous diffusion

Abstract

The ability of exogenous DNA cargo to overcome the active and viscoelastic eukaryotic cytoplasm is a principal determinant for the gene delivery efficacy. During DNA electrotransfer, DNA forms complexes with the membrane (DNA cargo) which is transported through the cytoplasm through a combination of passive diffusion and active transport. However, this process is poorly understood limiting rational optimization of DNA cargo to be delivered to different cell types. We have investigated the intracellular transport of DNA cargo (of sizes 100 bp, 250 bp and 500 bp) delivered by electrotransfer to non-cancerous and cancerous mammalian cells. We demonstrate that intracellular DNA cargo transport is governed by coexisting ergodic and non ergodic anomalous diffusion for all the tested DNA sizes and cell types. The apparent diffusion coefficient of the electrotransferred DNA cargo in the cytoplasm decreases when the DNA size is increased from 100 bp to 500 bp. Interestingly, the electrotransferred DNA cargo transport is strongly dependent on the cell's cancer state. Intracellular electrotransferred DNA cargo transport has a higher probability of superdiffusive transport in metastatic cells compared to malignant cells followed by benign cells.

Time-averaging and emerging nonergodicity upon resetting of fractional Brownian motion and heterogeneous diffusion processes

“…The model of FBM was recently applied to rationalize (•) the non-Brownian anomalous dynamics of lipids, cholesterols, and proteins in/on lipid membranes [155][156][157][158] , (•) the dynamics of G-proteins and G-protein-coupled receptors on plasma membrane 159,160 , (•) the diffusion of labeled mRNAs in living bacterial cells 161 , (•) anomalous motion of lipid granules in living yeast cells 162 , (•) the diffusion of telomeres inside the nuclei of human cancer cells 163,164 , (•) the dynamics of chromosomal loci in bacterial cells [165][166][167] , (•) non-Gaussian nonergodic anomalous diffusion of micron-sized beads in mucin-polymer hydrogels 168 , (•) tracer dynamics in actin networks 169 [for the particles larger in size than the network meshsize] 170 , (•) heterogeneous intracellular transport of DNA cargo in cancerous cells [with coexisting ergodic-andnonergodic but nonaging dynamics] 171 , as well as (•) intermittent bulk-surface non-Gaussian and aging anomalous diffusion with aging of anticancer-drug doxorubicin in silica nanoslits 172 , to mention a few examples.…”

Microtrap Array on a Chip for Localized Electroporation and Electro-Gene Transfection

“…In contrast to small molecules such as propidium iodide, larger charged plasmid DNA first forms DNA-membrane complexes upon electrotransfer and is later internalized through cellular machinery [51][52][53][54]. Since the intracellular release of plasmid DNA is difficult to control without the use of endosomal/nuclear disrupting or redirecting agents [55][56][57], the amount of plasmid DNA that forms the DNA membrane complex and the cell viability is key to successful cell transfection [58,59].…”

Activity-Induced Enhancement of Superdiffusive Transport in Bacterial Turbulence

“…This transport is also crucial for the survival of microorganisms that are thus able to dynamically regulate the living environment throughout all the scales. In order to break through the diffusion limit of random-work process, a variety of organism systems exploit anomalous diffusion, i.e., ballistic or superdiffusion to promote living activity such as quorum sensing [ 1 , 2 ], intracellular chromosome transport [ 3 ], cell migration [ 4 ], foraging and feeding [ 5 ], colony expansion [ 6 ], etc. In non-living chaotic flows, the swirls and jets cause the sticking and jumping processes of the immersed tracer particles, then make tracers diffuse much faster by the Lévy flight motion [ 7 , 8 ].…”