Elaboration of delivery system for DNA, RNA, protein, peptide, and smal molecule medical preparations
Funding: State Research Programme "Development of advanced prevention strategies, treatment, diagnostic tools and methods, biomedical technologies for improving public health":
Head of subproject: Dr. biol. Tatjana Kozlovska, Dr. chem. Regīna Renhofa
Dr. chem. Regīna Renhofa
Aim: To elaborate specific delivery systems, that provide targeted transport of DNA, RNA, proteins, peptides, small-molecular and inorganic materials to eukaryotic cells in vitro and in vivo.
Tasks: For 2010.- To develop chimeric or mosaic virus-like particles (VLPs), based on self-assembling bacteriophage GA coat protein (CP),that are equipped with addresses for specific interactions with cell cultures. Further – to investigate exploitation opportunities of such particles as delivery vectors and carriers of therapeutics via disassociation-reassociation-packaging mechanisms. To involve into investigations also VLPs based on Hepatitis B virus core-protein.
Results in 2010. : One-gene and two-gene constructions at all 26 were obtained. One-gene constructions were planned for expression of chimeric proteins where address sequences were N-terminally fused to GA CP sequence, but two-gene constructions – for co-expression of wild type GA CP and chimeric proteins. There were choosed three addresses – transducing HIV-1 TAT (14 amino acids), another transducing s.c. PTD4 (11 amino acids) and unspecific ligand for cell surface receptors-21 amino acid long N-terminal fragment of viral inflammatory protein vMIPII sequence. The fact has been proved, that elected addresses could be located only on mosaic VLPs, after studies of expression and VLP formation in E.coli and yeast Saccharomyces cerevisiae expression systems. For production and further investigation there were selected 4 constructions: one with TAT, one with PTD4 and two with vMIPII sequence.