Full-text resources of PSJD and other databases are now available in the new Library of Science.
Visit https://bibliotekanauki.pl
Preferences help
Polish version English version Language
enabled [disable] Abstract
Number of results

Results found: 5

Number of results on page
first rewind previous Page / 1 next fast forward last

Search results

Search:
in the keywords:  transfection
help Sort By:

help Limit search:
first rewind previous Page / 1 next fast forward last
1
Content available remote

New cationic polyprenyl derivative proposed as a lipofecting agent

100%
Madeja Z., Rak M., Wybieralska E., Różański I., Masnyk M., Chmielewski M., Łysek R., Chojnacki T., Jankowski W., Ciepichal E., Swiezewska E., Tekle M., Dallner G.
Acta Biochimica Polonica
|
2007
|
vol. 54
|
issue 4
873-876
EN
Cationic linear poly-cis-isoprenoid prepared from natural plant polyprenol in a mixture with dioleyl phosphatidylethanolamine was found to be an effective lipofection agent for eukaryotic cells. The transfecting activity is related to the poly-cis structure of the polyprenyl chain.
2
Content available remote

Electroporated intact BY-2 tobacco culture cells as a model of transient expression study.

100%
Kościańska E., Wypijewski K.
Acta Biochimica Polonica
|
2001
|
vol. 48
|
issue 3
657-661
EN
Transfer of foreign genes into plant cells can be accomplished by several methods: agrobacterium-mediated, microinjection, biolistic particle bombardment and electroporation. The last one is frequently used for transfection of plant protoplasts for transient gene expression. Electroporation is a simple procedure and allows transfecting a large number of cells at one time. Square wave-modulated porators are the most efficient for introducing expression cassettes into plant protoplasts. Based on a protocol developed by Wu & Feng (Plant Cell Reports, 1999, 18, 381-386), we optimized conditions for transfection of intact Nicotiana tabacum BY-2 cells using square wave-modulated electroporator. To simplify screening for transfected gene expression we used constructs with a GFP marker gene.Electroporation of cells in the presence of DNA has been widely used in recent years in molecular biology for studying transient gene expression. It consists in subjecting cells to an electric field, which forms pores in the lipid bilayer of the cell membrane, allowing DNA molecules to enter into the cytoplasm [1]. Pore formation is reversible and cell survival is maintained, thus such a method of introducing foreign DNA into cells is fast, simple, efficient, non-toxic and applicable to a great variety of cells. However, in spite of all its advantages electroporation has not been applied equally successfully in experiments with plant cells (except those with protoplasts) because of the cell wall. There are some earlier reports indicating that the cell wall does not prevent DNA molecules from being internalized [1-3]. In 1999 Wu and Feng [4] described an effective method of electroporation applicable to the intact plant cells. In this method plant cells are subjected to plasmolysis prior to electroporation. The modifications of these procedure are presented in this paper.
3
Content available remote

Optimisation of transfection conditions of CD34+ hematopoietic cells derived from human umbilical cord blood

100%
Ołdak T., Kruszewski M., Machaj E. K., Gajkowska A., Pojda Z.
Acta Biochimica Polonica
|
2002
|
vol. 49
|
issue 3
625-632
EN
Human umbilical cord blood is frequently used as a source of transplantable hematopoietic cells and more recently as a target of gene therapy - a new approach for treatment of various disorders. The aim of our study was optimisation of the transfection conditions of cord blood-derived CD34+ hematopoietic cells. Mononuclear cells fraction was isolated from cord blood samples by density gradient centrifugation. Subsequently, CD34+ hematopoietic cells were separated on immunomagnetic MiniMACS columns. Pure population of CD34+ cells was incubated in a serum free medium supplemented with thrombopoietin, stem cell factor and Flt-3 ligand for 48 h and then transfected with plasmid DNA carrying the enhanced version of green fluorescent protein (EGFP) as a reporter gene. We studied the influence of various pulse settings and DNA concentrations on the transfection efficiency, measured by flow cytometry as the fluorescence of target cells due to the expression of EGFP. The optimal settings were as follows: 4 mm cuvette, 1600 μF, 550 V/cm, and 10 μg of DNA per 500 μl. With these settings we obtained a high transfection frequency (41.2%) without a marked decrease of cell viability. An increase of the pulse capacitance and/or of DNA concentration resulted in a greater electroporation efficiency, but also in a decrease of cell viability. In conclusion, the results described here allow one to recommend electroporation as an efficient method of gene delivery into CD34+ hematopoietic cells derived from human umbilical cord blood.
4
Content available remote

Vehicles for Small Interfering RNA transfection: ExosomesversusSynthetic Nanocarriers

88%
Duechler M.
DNA and RNA Nanotechnology
|
2013
|
vol. 1
|
issue 1
EN
Therapies based on RNA interference (RNAi) hold a great potential for targeted interference of the expression of specific genes. Small-interfering RNAs (siRNA) and micro-RNAs interrupt protein synthesis by inducing the degradation of messenger RNAs or by blocking their translation. RNAibased therapies can modulate the expression of otherwise undruggable target proteins. Full exploitation of RNAi for medical purposes depends on efficient and safe methods for delivery of small RNAs to the target cells. Tremendous effort has gone into the development of synthetic carriers to meet all requirements for efficient delivery of nucleic acids into particular tissues. Recently, exosomes unveiled their function as a natural communication system which can be utilized for the transport of small RNAs into target cells. In this review, the capabilities of exosomes as delivery vehicles for small RNAs are compared to synthetic carrier systems. The step by step requirements for efficient transfection are considered: production of the vehicle, RNA loading, protection against degradation, lack of immunogenicity, targeting possibilities, cellular uptake, cytotoxicity, RNA release into the cytoplasm and gene silencing efficiency. An exosomebased siRNA delivery system shows many advantages over conventional transfection agents, however, some crucial issues need further optimization before broad clinical application can be realized.
5
Publication available in full text mode
Content available

Niewirusowe preparaty genowe – charakterystyka metod oceny właściwości fizykochemicznych

63%
Kuźnicka E., Małecki M.
Annales Academiae Medicae Silesiensis
|
2011
|
vol. 65
|
issue 5-6
67-79
EN
Gene therapy is based on the use of viral and non-viral nucleic acid preparations. They allow to administration of the therapeutic genes into targeted cells. Due to the lack of safe and eff ective methods of gene delivery, use of gene therapy in the clinic is restricted. The large attention is focused on studies of non-viral vectors, which seem to be stable and safe gene vehicles. Molecular complexes between plasmid DNA and cationic polymers, such as cationic lipids, polypeptides are formed spontaneously, as a result of electrostatic interactions. The mode of interaction of pDNA with polymeric compounds has a strict infl uence on physico-chemical properties of pDNA : cationic vector complexes. It is closely related to the gene transfer effi ciency (transfection). The evaluation of physical and molecular mechanisms of complex formation is crucial for effi cient gene administration into the cells. In this article various methods of studies of physicochemical properties of pDNA : vehicle complexes are described with special attention to their experimental/clinical value.
PL
Stanowiące podstawę terapii genowej geny kodujące terapeutyczne białka wprowadzane są do komórek za pomocą nośników wirusowych i niewirusowych. Brak jednoznacznie bezpiecznych, a jednocześnie efektywnych metod wprowadzania genów sprawia, iż zastosowanie terapii genowej w klinice jest wciąż ograniczone. Prowadzone są intensywne badania poświęcone wektorom niewirusowym – nośnikom genów, które wydają się stabilne in vitro i bezpieczne dla pacjentów. Kompleksy molekularne plazmidowego DNA (pDNA) i związków polimerowych tworzą się spontanicznie, w wyniku oddziaływań elektrostatycznych. Sposób oddziaływania pDNA ze związkami polimerowymi wpływa na właściwości fi zykochemiczne kompleksów pDNA : wektor kationowy, które mają ogromne znaczenie dla efektywności wprowadzania genów do komórek (transfekcji). Wdrażanie nowych, aktywnych biologicznie preparatów genowych do badań eksperymentalnych wiąże się z koniecznością prowadzenia prac poświęconych ocenie ich właściwości fi zykochemicznych i biochemicznych, które wykonuje się metodami stosowanymi w laboratoriach inżynierii genetycznej; często szuka się również nowych rozwiązań analitycznych. W artykule podjęto próbę scharakteryzowania niewirusowych preparatów genowych głównie w kontekście oceny ich właściwości fizykochemicznych. Opisano aktualnie stosowane metody badania preparatów genowych. Świadomość możliwości prowadzenia w pracowniach różnych dyscyplin naukowych badań właściwości kompleksów pDNA : : nośnik może pomóc w projektowaniu nowych preparatów genowych i próbach ich wykorzystania w leczeniu chorych.
first rewind previous Page / 1 next fast forward last
JavaScript is turned off in your web browser. Turn it on to take full advantage of this site, then refresh the page.