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.
Tang
Yizhe
Y
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.
Le
Long P
LP
Matthews
Qiana L
QL
Han
Tie
T
Wu
Hongju
H
Curiel
David T
DT
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