ISSN 1995-5537
«Біотехнологія» journal V. 5, No. 5, 2012
Р. 16-26, Bibliography 73, English.
Universal Decimal classification: 602.641:578.826 602.64 602.9
ADENOVIRAL VECTORS: CONVENIENT TOOLS FOR GENE DELIVERY TO PRIMARY MAMMALIAN CELLS
Institute of Cell Biology of National Academy of Sciences of Ukraine, Lviv
University of North Carolina at Chapel Hill, Chapel Hill, NC, USA
Virginia Commonwealth University, Richmond, VA, USA
Tissue cell culture is widely used as convenient technique in modern biology and drug discovery. Multiple cell lines derived from normal and neoplastic tissues are used world-wide. However all the established cell lines, including pseudo-normal ones, are at least minimally transformed and frequently cannot properly re-capitulate the in vivo situation. Many of these limitations can be circumvented by using primary cells. Unfortunately, these cells are frequently difficult to transfect. The E1/E3-deficient recombinant adenoviruses shown to be highly efficient gene delivery tools that allow successful work with multiple types of difficult to transfect cells in different in vivo applications. Generation of the recombinant adenoviral constructs includes sub-cloning of the insert into a shuttle vector with convenient polycloning site with subsequent homologous recombination of the shuttle vector with an adenoviral backbone. We described a generation of more than 80 new rAdvs including expression, luciferase reporter and small hairpin interfering RNA vectors by using the AdEasy system. The generated recombinant adenoviruses can be used in multiple biological applications and in combination with proper maintenance protocol allow for 100% efficiency of transduction of difficult to transfect cells including primary mouse keratinocytes, human bone marrow-derived primary mesenchymal stem cells and other types of primary mammalian cells.
Key words: adenoviral vector; transduction, primary mammalian cells, mesenchymal stem cells.
© Palladin Institute of Biochemistry of National Academy of Sciences of Ukraine, 2008