Projects / Programmes
Elektroporacija kot metoda za vnos funkcionalnih membranskih proteinov v celice sesalcev brez uporabe genske manipulacije (Slovene)
| Code |
Science |
Field |
Subfield |
| 2.06.07 |
Engineering sciences and technologies |
Systems and cybernetics |
Biomedical technics |
| Code |
Science |
Field |
| 2.06 |
Engineering and Technology |
Medical engineering
|
Researchers (8)
Organisations (1)
Significance for science
Within the framework of the project, we showed for the first time that polymersomes (PS) can also be porated with nanosecond electric pulses. The advantage of these pulses is that their effect on the poration is less dependent on the size of the vesicles and cells and could allow the fusion of structures of different sizes. We also found that the PS are porating at high amplitudes, duration and number of electric pulses, which indicates their stability and rigidity. We also investigated the effect of the repetition frequency of electrical pulses on PS deformation in different solutions (inside the PS and outside) and found that under certain conditions they extend in the direction of the electric field, which is an advantage in providing contacts for possible fusion with cells and other structures. We compared the phenomena with similar deformations of lipid vesicles (liposomes) under the influence of alternating current. We also found that PS deformations are more prolonged than in liposomes, and that they do not affect the poration, which further indicates the high PS stability. This probably contributed significantly to the fact that we were not able to fuse them with cells. We have also shown that PS in different forms (large, giant unilamellar vesicles) are not toxic to two types of cells (CHO, B16F1, up to 250 µg / ml for 48 h or 1.35 mg / ml for 60 min). This means that these PS are suitable for the research of various interactions with mammalian cells. A review of the methods of detection of electroporation (In vitro electroporation detection methods: an overview) received an exceptional response from Slovenian and foreign collaborators, and we believe that it will serve its purpose well, namely to make it easier for researchers to choose the appropriate method for detecting electroporation in vitro, which will contribute to the faster progress of this scientific field.
Significance for the country
Within the framework of the project, basic investigations of poration and deformation of polymersomes, synthetic vesicles from amphiphilic copolymers, under the influence of nanosecond electric pulses and the establishment of contacts between cells and polymersomes (PS) with dielectrophoresis were carried out. We also found that these PS are not toxic to two types of mammalian cells and could serve as biocompatible carriers of proteins. This has laid the foundations for fusion of cells and PS as a new method for delivering transmembrane proteins to the cell membrane without the use of gene transfection. Unfortunately, we were not able to set the method. However, we hope that this research is the basis for the establishment of this method, which could, as an alternative to gene transfection, have a significant effect on the treatment of hereditary diseases associated with defective membrane protein genes. By connecting with a foreign partner group, we strengthened the involvement of Slovenian researchers in the international scientific network, which is important for the development of Slovenian science. In doing so, we gained new contacts for the design of new scientific projects, with which we applied for international funding (for example, for European funds). By doing this, we try to provide jobs for Slovenian researchers.
Most important scientific results
Annual report
2015,
final report
Most important socioeconomically and culturally relevant results
Annual report
2015,
final report
