Engineering novel cell surface receptors for virus-mediated gene transfer.
The absence of viral receptors is a major barrier to efficient gene transfer in many cells. To overcome this barrier, we developed an artificial receptor based on expression of a novel sugar. We fed cells an unnatural monosaccharide, a modified mannosamine that replaced the acetyl group with a levulinate group (ManLev). ManLev was metabolized and incorporated into cell-surface glycoconjugates. The synthetic sugar decorated the cell surface with a unique ketone group that served as a foundation on which we built an adenovirus receptor by covalently binding biotin hydrazide to the ketone. The artificial receptor enhanced adenoviral vector binding and gene transfer to cells that are relatively resistant to adenovirus infection. These data are the first to suggest the feasibility of a strategy that improves the efficiency of gene transfer by using the biosynthetic machinery of the cell to engineer novel sugars on the cell surface.
Howard Hughes Medical Institute and Departments of Internal Medicine and Physiology and Biophysics, University of Iowa, Iowa City, Iowa 52242, USA. mjwelsh@blue.weeg.uiowa.edu
J H
JH
Lee
T J
TJ
Baker
L K
LK
Mahal
J
J
Zabner
C R
CR
Bertozzi
D F
DF
Wiemer
M J
MJ
Welsh
D016475:3T3 Cells; D000256:Adenoviridae; D000818:Animals; D001710:Biotin / Q000031:analogs & derivatives / Q000493:pharmacokinetics; D002478:Cells, Cultured; D004730:Endothelium, Vascular / Q000166:cytology / Q000502:physiology*; D018014:Gene Transfer Techniques; D017930:Genes, Reporter; D005822:Genetic Vectors; D006595:Hexosamines / Q000493:pharmacokinetics*; D006801:Humans; D007700:Kinetics; D051379:Mice; D008954:Models, Biological; D011956:Receptors, Cell Surface / Q000096:biosynthesis / Q000502:physiology*; D019809:Streptavidin / Q000493:pharmacokinetics; D014162:Transfection / Q000379:methods*; D014471:Umbilical Veins; D001616:beta-Galactosidase / Q000096:biosynthesis / Q000235:genetics*
D006595:Hexosamines; C105871:N-levulinoyl mannosamine; D011956:Receptors, Cell Surface; C053311:biotin hydrazide; D001710:Biotin; D019809:Streptavidin; D001616:beta-Galactosidase