ABSTRACT
The patterns of formation of RNA nanoparticles (NPs) during thermal cycling of bacterial total tRNA in the presence of cations Ca2+, Mn2+, Ni2+, Zn2+, Co2+, and Cu2+ were studied. The optimal conditions for the production of NPs were found, and it was revealed that their size depends on the ratio of the concentrations of Me2+ and tRNA. The concentration of reagents for obtaining NPs of small size (from 5 to 100 nm) was selected. It was shown that tRNA-based nanoparticles can comprise short (20-50 nt) ribooligonucleotides, including aptamers and siRNAs. The stability of NPs during storage in buffer solutions of various composition was studied. It was found that the initial suspensions of NPs are quite stable, but they are rapidly destroyed in PBS buffer (pH 7.4). A simple and effective stabilizer (polyarginine) was found, the additives of which ensure the preservation of nanoparticles in PBS buffer for more than 5 h. Nanoparticles modified with the stabilizer are resistant to blood serum nucleases and can be used for transfection.
Subject(s)
Cations, Divalent/chemistry , Nanoparticles/chemistry , RNA, Transfer/chemistry , Particle SizeABSTRACT
Histone H2A can deliver transgenic DNA into mammalian cells in vitro. The ability of DNA delivery in vivo is a question of the further work, but it is possible to estimate factors in in vitro experiments, which affect delivery in vivo. The first step in this direction was to determine sizes and ζ-potentials of histone H2A complexes with DNA. In this work, we produced recombinant histone H2A and its modification, containing TAT-peptide from human immunodeficiency virus TAT protein, which is capable of enhancing macromolecule delivery into mammalian cells. The effective diameters and ζ-potentials of histones-DNA complexes were estimated. Complexes of histone H2A and complexes of modified histone H2A-TAT with DNA had positive ζ-potentials. Complexes of histone H2A with DNA had an effective diameter of about 200 nm. Histone modification with TAT-peptide led to aggregation and formation of massive particles of about 1 µm in diameter.
Subject(s)
DNA/chemistry , Histones/chemistry , tat Gene Products, Human Immunodeficiency Virus/chemistry , HIV-1/chemistry , HIV-1/genetics , Histones/genetics , Humans , Particle Size , Plasmids/chemistry , Plasmids/genetics , tat Gene Products, Human Immunodeficiency Virus/geneticsABSTRACT
The effects of 30-min medium-intensity exercise on the expression of genes encoding heat shock protein 70 (HSPA1A) and its cochaperones HSP-70-binding protein 1 (HSPBP1) and Tag7 (PGLYRP1) in human leukocytes were studied. Transcription activities of HSPA1A and PGLYRP1 genes increased immediately after medium-intensity exercise, while activity of HSPBP1 gene remained unchanged. During recovery after exercise, the expression of HSPA1A gene virtually did not change, while the expression of PGLYRP1 gene continued to increase and after 90 min more than 2-fold surpassed the basal level.
