Derivation of a triple mosaic adenovirus based on modification of the minor capsid protein IX.
Adenoviral capsid protein IX (pIX) has been shown to be a potential locale to insert targeting, imaging-related and therapeutic modalities by genetic modification. Recent evidences suggested that capsid protein mosaicism could be a promising strategy for improving the utility of Ad vector. In this study, we explored a method to genetically generate triple pIX mosaic Ad serotype 5 (Ad5) displaying three types of pIX on a single virion. pIXs were modified at their carboxy termini with a Flag sequence, a hexahistidine sequence (His(6)) or a monomeric red fluorescent protein (mRFP1), respectively. Western blotting analysis and fluorescence microscopy of the purified recombinant viruses indicated that all three modified pIXs were incorporated into the viral particles. Immuno-gold electron microscopy (EM) further confirmed that three types of pIX indeed co-existed on an individual virion. These results firstly validated a triple mosaic capsid configuration on pIX, and demonstrated the possibility of further radical design.
Division of Human Gene Therapy, Department of Medicine, and the Gene Therapy Center, University of Alabama at Birmingham, 901 19th Street South, BMR2-502, Birmingham, AL 35294, USA.
Yizhe
Y
Tang
Long P
LP
Le
Qiana L
QL
Matthews
Tie
T
Han
Hongju
H
Wu
David T
DT
Curiel
D000260:Adenoviruses, Human / Q000235:genetics* / Q000502:physiology; D000956:Antigens, Viral / Q000235:genetics / Q000378:metabolism; D002213:Capsid / Q000737:chemistry*; D036022:Capsid Proteins / Q000235:genetics*; D005818:Genetic Engineering / Q000379:methods; D005822:Genetic Vectors; D008856:Microscopy, Fluorescence; D010455:Peptides / Q000737:chemistry* / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D014771:Virion / Q000648:ultrastructure*
D000956:Antigens, Viral; D036022:Capsid Proteins; D010455:Peptides; D011993:Recombinant Fusion Proteins