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Content archived on 2024-06-18

Single Cell Electroporation and DNA dynamics: from bulk to micro/nanofluidics

Objective

Electroporation or electropermeabilization is the electrical disruption of a cell’s membrane to introduce foreign DNA, RNAs, drugs, proteins, or other therapies into the living cells. This technique is one of the most popular non-viral gene transfer methods for both in vitro and in vivo application. However, this method is suffering from several limitations such as: i) low transfection efficiency, ii) poor cell viability, and iii) random uptake. Multiple applications of electroporation in biotechnology and medicine rely on trial and error optimization of treatment conditions rather than knowledge of pores, dynamics and electrotransfer of DNA, due to the limited spatial and temporal resolution of traditional tools. More interesting, the phenomena behind electro-mediated membrane permeabilization to plasmid DNA (~4.5 kbp) have been shown to be significantly more complex than those for small molecules. Therefore, successful DNA electrotransfer into cells depends not only on cell permeabilization (or poration) but also on the way plasmid DNA interacts with the membrane and, once into the cell, migrates toward the nuclei.

The main aim of this proposal to understand and control the transport of DNA in electroporation treatment such that stable, safe and efficient gene transfection can be achieved. Here I will use single molecule techniques (such as AFM, FRET, confocal and optical tweerers) with high spatio-temporal resolution to investigate the process of electroporation at different length (from bulk to micro/nano) and time scale (ranging from micro-seconds to hours). In addition, these unique tools will be integrated with novel micro/nanofluidics to investigate the electroporation and electrokinetic flow of DNA at micro/nano-scale. I will observe all the processes in real time with unprecedented details and quantitatively examine the kinetics of pore formation/resealing and cytoplasmic trafficking of DNA.

Fields of science (EuroSciVoc)

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Topic(s)

Calls for proposals are divided into topics. A topic defines a specific subject or area for which applicants can submit proposals. The description of a topic comprises its specific scope and the expected impact of the funded project.

Call for proposal

Procedure for inviting applicants to submit project proposals, with the aim of receiving EU funding.

FP7-PEOPLE-2012-CIG
See other projects for this call

Funding Scheme

Funding scheme (or “Type of Action”) inside a programme with common features. It specifies: the scope of what is funded; the reimbursement rate; specific evaluation criteria to qualify for funding; and the use of simplified forms of costs like lump sums.

MC-CIG - Support for training and career development of researcher (CIG)

Coordinator

TECHNISCHE UNIVERSITEIT DELFT
EU contribution
€ 100 000,00
Total cost

The total costs incurred by this organisation to participate in the project, including direct and indirect costs. This amount is a subset of the overall project budget.

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