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1

Genetically engineered potato resistant to potato leafroll virus infection

100%
Palucha A., Chrzanowska M., Zagorski W., Hulanicka D.
Biotechnologia
|
1997
|
issue 4
38-47
EN
Binary plasmids carrying different fragments of the coat protein gene of potato leafroll luteovirus were constructed for transformation of commercial potato cultivar Bzura. Several transgenic plant lines were obtained and characterised. The resistance of transgenic plants to the virus was tested by inoculation with aphids or grafting. Selected transformants expressing viral RNA were resistant to virus challenge by viruliferous aphids. Expression of the antisense RNA prevented virus infection even after grafting with scions from infected plants.
2

Oligonucleotide conjugates

100%
Misiura K.
Biotechnologia
|
1994
|
issue 4
55-58
EN
Various molecules are attached to oligonucleotides to make conjugates.Conjugates are specially designed to increase antisense activity by better interaction with target mRNA, better stability against nucleases, and/or permeability to the cells.
3

Background of antisense strategy and potential applications

80%
Madej-Dudzinska B., Twardowski T.
Biotechnologia
|
1999
|
issue 3
62-75
EN
Structure and function of antisense oligomers and ribozynes are discussed. Some applications of aDNA in medicine and agrobiotechnology are discussed.
4

Mechanistic studies of Hammerhead ribozytmes and their application in vivo

80%
Zhou D.-M., Kohda T., Koseki S., Yamamoto R., Yasugi M., Tsaguawa K., Ohkawa J., Koguma T., Taira K.
Biotechnologia
|
1996
|
issue 4
183-200
EN
The hammerhead ribozyme belongs to the class of molecules known as antisense RNAs.However, because of short extra sequences that form the so-called catalytic loop, it can act as an enzyme.Since the catalytic domain captures magnesium ions and magnesium ions can cleave phosphodiester bonds, hammerhead ribozymes are recognized as metalleozymes.In RNA cleaving reaction catalyzed by protein enzymes, the cleavage of phosphodiester bonds involves acid/base catalysis, with proton transfer occurring in the transition solvent isotope effects, in reaction catalyzed by hammerhead ribozymes, it became apparent that no proton transfer occurs in the transition state during reactions catalysed by a hammerhead ribozyme.This and an additional kinetic analysis, using a natural all-RNA substrate that contains a 5'-thio-leaving group at the cleavage site, revealed that hammerhead ribozymes exploit the general double-metal-ion mechanism of catalysis, with Mg2+ ions coordinating directly with the attacking and leaving oxygen moities.Since the hammerhead ribozyme is one of the smallest RNA enzymes known and has potential as a antiviral agent, thus ribozyme has been extensively investigated for application in vivo.Ribozymes are described that have possible utility as agents against HIV-1.
5

Strategy of directed RNA degradation and selected examples of its use in antiviral therapy

80%
Dutkiewicz M., Ciesiolka J.
Biotechnologia
|
2002
|
issue 1
57-70
EN
Over the last decade, antisense oligodeoxynucleotides and ribozymes have emerged as valuable biochemical tools and promising therapeutic reagents in medicine. This review describes the basic principles of their use in the strategy of directed RNA degradation. The rules for the rational choice of targeted RNA sequences, the properties of available RNA cleaving tools, as well as the major problems that limit the effectiveness of this new technology, are discussed. Selected examples of the successful use of antisense oligonucleotides and ribozymes as antiviral agents are presented.
6

Interference in ethylene biosynthetic pathway using transformation

70%
Kepczynski J., Kepczynska E.
Biotechnologia
|
2000
|
issue 2
72-82
EN
The biosynthesis of ethylene in plants and its regulation by manipulating the expression of ACC synthase or ACC oxidase genes are discussed. Ethylene synthesis can be reduced by the introduction of antisense ACC synthase or antisense ACC oxidase genes. Expression genes of SAM hydrolase from bacteriofage T3, which catalyze the conversion of SAM to methylothioadenosine, also diminished ACC availability. Another possibility of ethylene biosynthesis control is the expression of gene encoding ACC deaminase from Pseudomonas.
7

From neoplastic neural development to gene therapy of brain tumors ? IGF-I antisense and triple helix approaches

70%
Trojan J.
Biotechnologia
|
2003
|
issue 2
280-289
EN
IGF-I, insulin ? like growth factor I, seems to play a major role in the normal and tumoral development of the nervous system. Glioblastoma is the most frequent brain tumor in man and is usually fatal. Both human and rat glioma cells express high amounts of IGF-I. When rat glioma cells are transfected with vectors expressing either IGF-I antisense RNA or inducing IGF RNA ? DNA triple helix, the synthesis of IGF-I was stopped on translation or transcription levels, respectively. Down-regulation in the expression of IGF-I coincides with the reappearance of B-7 and MHC class I antigens at the surface of transfected cells. When injected subcutaneously, the transfected cancer cells initiate an immune reaction involving CD8+ lymphocytes, followed by tumor regression. The ?anti-gene? strategy for clinical therapy of glioblastoma, and other tumors expressing IGF-I such hepatomas were introduced in University Hospitals of Cleveland (USA), Shanghai ( China) , Krakow and Bydgoszcz (Poland).
8

In vitro regeneration and transformation of rye (Secale cereale L.)

70%
Sowa S., Sowa A., Zimny J.
Biotechnologia
|
2000
|
issue 4
88-92
EN
In vitro cultures are an integral part of plant transformation. Genetic manipulation can be performed only on a single cell level. Therefore in vitro culture and regeneration of plants from a single cell are very important for successful transformation In vitro culture of rye is more difficult to conduct than of others cereals. Difficulties with in vitro regeneration of rye seems to be the main factor limiting the development of rye transformation systems. Genetic transformation process includes three main steps, single cell transformation, selection of transgenic cells and regeneration of plants from single cells. Efficiency of each of these steps can influence the result of the transformation process. Therefore optimisation of those steps is very important.Transformation has been performed using the microprojecticle method and scutellum of rye embryos as a target. Two constructs have been used in cotransformation, pDB1 containing a marker bar gene (phosphinotricine acetylotransferase) for Basta herbicide resistance and a repoter uidA gen (beta-glucuronidase) and pAwact-Sec containing 196 bp fragment of a sec-1 gene in anti-sense orientation. Using pAWact-Sec construct we tried to block the expression of the endogenous gene sec-1 coding omega-secalin which is storage protein of rye grain. We were able to regenerate transgenic rye plants containing all introduced genes. The efficiency of sec-1 expression blocking was analysed by SDS-PAGE method. Among 50 analysed kernels of T1 generation we found 5 with lower omega-secalin level. Complete blocking of omega-secalin was not observed.
9

Oligonucleotide transport across the plasma membrane

61%
Laski J.
Biotechnologia
|
1994
|
issue 4
40-49
EN
Transport of synthetic oligonucleotides across the plasma membrane of mammlian cells is the one of the major problems in the application of the antisense strategy.Modifications of oligonucleotides may improve their transport and stability in the cell's medium.The therapeutical agents tergeted to intracellular structures and their mechanisms are reviewed.
10
Content available remote

Effect of opioids on Ca2+/cAMP responsive element binding protein

61%
Bilecki W., Przewlocki P. R., Przewlocki R.
Acta Neurobiologiae Experimentalis
|
2000
|
vol. 60
|
issue 4
557-567
EN
Ca2+/cAMP response element binding protein (CREB) is an important factor linking the opioid-regulated secondary messenger systems to alterations in gene expression. Opioids regulate CREB level, its phosphorylation and binding to its corresponding response element in the promoters of several genes implicated in drug addiction. CREB mediates the action of opioids on the expression of several genes in brain regions responsible for drug-seeking behavior and manifestation of signs of dependence. Moreover, alterations in CREB level can affect the rewarding properties of morphine and regulate the self-administration of cocaine. At the cellular level CREB acts as convergence point for different cellular pathways. Opioids affect two different intracellular mediator systems: inhibitory - connected with cAMP, and stimulatory - involving calcium and the PKC pathway. Both can affect CREB but in different phases of opiate action. The presence of this biphasic mechanism can explain the phenomenon of the induction of some CRE-controlled genes after both acute and chronic morphine administration. Cellular studies also highlight the relevance of other ATF/CREB family members which can affect Ca2+/cAMP response element (CRE) controlled transcription as well as other transcription factors which make the opioid induction longer lasting.
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