ABSTRACT
We compared the results of real-time PCR analysis of gene expression in paired specimens breast cancer tissue fixed in RNAlater (Qiagen) stabilization reagent (FF samples) and in formalin (FFPE samples). A clear-cut linear relationship (correlation coefficient 0.76 ± 0.07) was detected between the gene expression logarithm values in FF and FFPE samples. This fact suggests that collections of paraffin blocks with formalin-fixed tissue specimens from patients with a many-year disease history can be effectively used in modern studies.
Subject(s)
Breast Neoplasms/metabolism , Carcinoma, Ductal, Breast/metabolism , Gene Expression Profiling/methods , Tissue Fixation/methods , Breast Neoplasms/genetics , Breast Neoplasms/pathology , Carcinoma, Ductal, Breast/genetics , Carcinoma, Ductal, Breast/pathology , Female , Fixatives/chemistry , Formaldehyde/chemistry , Gene Expression , Humans , Middle Aged , RNA Stability , Reagent Kits, DiagnosticABSTRACT
The ability of retroviruses and transposases to insert own genome into a host-cell allow us to consider them as a preferable object for constructing gene therapy vectors. However, enzymes that perform the insertion of the genetic material do not display a selectivity towards target nucleotide sequences that results in an almost random DNA introduction into the recipient cell genome. Random insertion leads to mutations which might cause a number of undesirable consequences including neoplastic transformation in the cell. Thereby, in order to achieve a successful functioning of retroviral and trasposonal genetic therapy systems, it is essential to modify them in such a way that directed integration of the vector in a target sequence in the human genome could be achieved. In the review approaches that have been developed for a high specific modification of genome, including the construction of hybrid proteins on the basis of retroviral integrases, transposases, as well as cellular factors interacting with these enzymes, are presented.
Subject(s)
Genetic Engineering/methods , Genetic Vectors/genetics , Integrases/metabolism , Retroviridae/genetics , Transposases/metabolism , Virus Integration/genetics , Adaptor Proteins, Signal Transducing/genetics , Genetic Therapy , Genome, Human , HIV-1/genetics , Humans , Integrases/genetics , Mutagenesis, Insertional , Transcription Factors/genetics , Transposases/geneticsABSTRACT
The binding of Gd3+ ions to linear double-stranded DNA molecules in water-salt solutions or in liquid-crystalline dispersions is accompanied by sharp changes in their optical and X-ray characteristics. Depending on the initial conditions of complex formation, the binding of Gd3+ ions either to DNA bases or phosphate groups occurs, which leads to changes in the properties of the liquid-crystalline dispersions. The packing of neighboring DNA molecules in particles of the liquid-crystalline dispersion of the complex DNA-Gd3+ depends strongly on the concentration of Gd3+ ions. This process is accompanied by a decrease in the amplitude of Bragg's reflection maximum. The unique properties of the developed material open the possibilities for its practical use.
Subject(s)
DNA/chemistry , Gadolinium/chemistry , Liquid Crystals/chemistry , Crystallography, X-Ray , Nucleic Acid Conformation , Solutions/chemistryABSTRACT
Human immunodeficiency virus type 1 integrase is one of the most attractive targets for the development of anti-HIV-1 inhibitors. The capacity of a series of 2,1,3-benzoxadiazoles (benzofurazans) and their N-oxides (benzofuroxans) selected using the PASS software to inhibit the catalytic activity of HIV-1 integrase was studied in the present work. Only the nitro-derivatives of these compounds were found to display inhibitory activity. The study of the mechanism of inhibition by nitro-benzofurazans/benzofuroxans showed that they impede the substrate DNA binding at the integrase active site. These inhibitors were also active against integrase mutants resistant to raltegravir, which is the first HIV-1 integrase inhibitor approved for clinical use. The comparison of computer-aided estimations of the pharmacodynamic and pharmacokinetic properties of the compounds studied and raltegravir led us to conclude that these compounds show promise and need to be further studied as potential HIV-1 integrase inhibitors.
ABSTRACT
Due to their ability to integrate into the host cell's genome, retroviruses represent an optimal basis for the creation of gene therapy vectors. The integration reaction is carried out by a viral enzyme integrase: thus, a detailed research of this enzyme is required. In this work, the catalytic properties of human foamy virus integrase were studied. This virus belongs to the Retroviridae family. The dissociation constant was determined, together with the kinetics of integrase catalytic activity. The data obtained were compared to those for the human immunodeficiency virus integrase and a considerable similarity in the activity of the two enzymes was observed.