Electrotransformation pathways of procaryotic and eucaryotic cells: recent developments

Eynard, Nathalie; Rols, Marie-Pierre; Ganeva, Valentina; Galutzov, B.; Sabri, N.; Teissié, Justin

doi:10.1016/s0302-4598(97)00038-x

Cited by 35 publications

(32 citation statements)

References 37 publications

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“…Plasmid/membrane complex remains accessible to DNase I, up to 60 s after the pulsation in the case of CHO cells. The plasmid aggregates, which are anchored in the membrane after the electric field application, remain sensitive to the degrading action of the nucleases that were added externally post-pulse, which are known not to cross the membrane (Eynard et al 1997). Nevertheless, the opposite observation, reported as a protective effect against DNAse, was detected when using a different protocol (Klenchin et al 1991).…”

Section: Biophysical Mechanismsmentioning

confidence: 98%

“…This process brings new properties to the cell plasma membrane, which, besides being permeabilized, becomes fusogenic and allows exogenous proteins to be inserted in it. Electropermeabilization is used to introduce a large variety of molecules into many different cells in vitro (Orlowski and Mir 1993;Eynard et al 1997). Clinical applications of the method are now under development as results of the EU Cliniporator and ESOPE programs.…”

Section: Introductionmentioning

confidence: 99%

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Gene electrotransfer: from biophysical mechanisms to in vivo applications

et al. 2009

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Electropulsation is one of the nonviral methods successfully used to deliver genes into living cells in vitro and in vivo. This approach shows promise in the field of gene and cellular therapies. The present review focuses on the processes supporting gene electrotransfer in vitro. In the first part, we will report the events occurring before, during, and after pulse application in the specific field of plasmid DNA electrotransfer at the cell level. A critical discussion of the present theoretical considerations about membrane electropermeabilization and the transient structures involved in the plasmid uptake follows in a second part.

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Section: Biophysical Mechanismsmentioning

confidence: 98%

Section: Introductionmentioning

confidence: 99%

Gene electrotransfer: from biophysical mechanisms to in vivo applications

et al. 2009

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“…τ load , is given by τ load = rC m (1/2λ out + 1/λ cyt ) [3] C m is the membrane capacitance, λ out and λ cyt , respectively, the conductance of the external buffer and of the cytoplasm. τ load is longer for larger cells in a heterogeneous population.…”

Section: Theory Of Membrane Potential Difference Modulationmentioning

confidence: 99%

“…66 Cell electroassociated DNA remains accessible to DNAaseI, a double-strand nucleic acid degrading enzyme, up to 60 s after the pulsation in the case of CHO cells. 3 The DNA aggregates, which are anchored in the membrane after the electric field application, remain sensitive to the degrading action of the externally post pulse added nucleases, which are known not to cross the membrane…”

Section: Events After Electropulsationmentioning

confidence: 99%

“…It has been used to introduce a large variety of molecules into many different cells in vitro. 2,3 Clinical applications of the electropermeabilization are now under development as a results of the EU Cliniporator and Esope programs. A local antitumoral drug delivery to patients (a method called electrochemotherapy) is under clinical trial.…”

Section: Introductionmentioning

confidence: 99%

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Time dependence of electric field effects on cell membranes. A review for a critical selection of pulse duration for therapeutical applications

Teissié¹,

Escoffre²,

Rols³

et al. 2008

Radiology and Oncology

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Background. Electropulsation is one of the non-viral methods successfully used to transfer drugs and genes into living cells in vitro as in vivo. This approach shows promise in field of gene and cellular therapies. This presentation first describes the temporal factors controlling electropermeabilization to small molecules (< 4kDa) and then the processes supporting DNA transfer in vitro. The description of in vitro events brings our attention on the processes occurring before (s), during (ms) and after electropulsation (ms to hours) ofhas the main advantages of being easy to use, fast, reproducible and safe.While during 30 years due to technological limits, pulse duration was always larger than 1 microsecond, the recent availability of high voltage (tens of kV) nanosecond long pulse generators opens the way to a new approach. Very fast perturbations under strong fields are induced in the membrane organization. 16,17 A new field of development is now present for electropermeabilization and promising results for clinical applications were reported.One of the limiting problems remains that very few is known on the physicochemical mechanisms supporting the reorganisation of the cell membrane. The molecular target of the field effect remains unclear.The present review focuses on the critical role played by the pulse duration in the electropermeabilization to small molecules (< 4kDa) and on its support to the processes associated to DNA transfer in vitro. Pulse durations are easy to adjust for an optimization of the clinical target: electrochemotherapy, irreversible electropermeabilization or gene therapy as suggested as a final conclusion. Electropermeabilization Theory of membrane potential difference modulation.An external electric field modulates the membrane potential difference. 18 From the physical point of view, a cell can be described as a spherical capacitor which is charged by the external electrical field. The transmembrane potential difference induced by the electric field, ΔΨi is a complex function g(λ) of the specific conductivities of the membrane (λ m ), the pulsing buffer (λ out ) and the cytoplasm (λ cyt ), the membrane thickness and the cell size. Thus, 1 ΔΨi = f. g (λ). r. E.cosθ [1] in which θ designates the angle between the direction of the normal to the membrane at the considered point on the cell surface and the field direction, E the field intensity, r the radius of the cell and f, which is a shape factor (a cell being a spheroid). Therefore, ΔΨi is not uniform on the cell surface. It is maximum at the positions of the cell facing the electrodes. These physical predictions were checked experimentally by videomicroscopy by using the potential difference sensitive fluorescent probes. [19][20][21] The pulse duration plays a critical role when shorter than the capacitive loading time of the membrane. In the previous part of the paper, it was considered that the pulse was long enough to bring the potential steady state value. The loading time τ load brings a limit in this description. 1 ΔΨi = f. ...

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Electro‐mediated gene transfer and expression are controlled by the life‐time of DNA/membrane complex formation

Faurie

Reberšek

Golzio

et al. 2009

The Journal of Gene Medicine

105

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Background Electroporation is a physical method used to transfer molecules into cells and tissues. Clinical applications have been developed for antitumor drug delivery. Clinical trials of gene electrotransfer are under investigation. However, knowledge about how DNA enters cells is not complete. By contrast to small molecules that have direct access to the cytoplasm, DNA forms a long lived complex with the plasma membrane and is transferred into the cytoplasm with a considerable delay.

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Electrotransformation pathways of procaryotic and eucaryotic cells: recent developments

Cited by 35 publications

References 37 publications

Gene electrotransfer: from biophysical mechanisms to in vivo applications

Gene electrotransfer: from biophysical mechanisms to in vivo applications

Time dependence of electric field effects on cell membranes. A review for a critical selection of pulse duration for therapeutical applications

Electro‐mediated gene transfer and expression are controlled by the life‐time of DNA/membrane complex formation

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