Protein-nucleic acid hybrid vector capable of easily binding genes

2014 Fiscal Year Final Research Report

• Project/Area Number: 24560955
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Biofunction/Bioprocess
• Research Institution: Akita University
• Principal Investigator: GOTOH Takeshi 秋田大学, 工学(系)研究科(研究院), 教授 (10215494)
• Project Period (FY): 2012-04-01 – 2015-03-31
• Keywords: gene delivery / streptavidin / TAT / NLS / insect cell

Outline of Final Research Achievements

Using cell penetrating peptide TAT (YGRKKRRQRR) from HIV-1 and nuclear localization signal NLS (PKKKRKV) from SV40 virus, which were attached to enhanced green fluorescent protein (EGFP), we examined the cellular uptake and nuclear localization of EGFP by Sf9 insect cells by confocal laser scanning microscopy. As a result, it was found that EGFP carrying both TAT and NLS at the N-terminal end in this order possessed the ability to cross the plasma membrane and nuclear membrane of Sf9 within 30 min. The 1:3 tetramer of active and inactive streptavidin (STVa and STVd), which had TAT and NLS at the N-terminal end in this order, was prepared, and a novel gene vector made up of the streptavidin tetramer and biotinylated gene was constructed. The biotinylated gene was prepared by annealing and ligating a synthetic biotinylated oligonucleotide and a gene having the complementary cohesive end. We are now investigating cellular uptake and nuclear localization of the vector by Sf9.

Free Research Field

生物化学工学