title: Development of a novel efficient method to construct an adenovirus library displaying random peptides on the fiber knob.
abstract: Redirection of adenovirus vectors by engineering the capsid-coding region has shown limited success because proper targeting ligands are generally unknown. To overcome this limitation, we constructed an adenovirus library displaying random peptides on the fiber knob, and its screening led to successful selections of several particular targeted vectors. In the previous library construction method, the full length of an adenoviral genome was generated by a Cre-lox mediated in vitro recombination between a fiber-modified plasmid library and the enzyme-digested adenoviral DNA/terminal protein complex (DNA-TPC) before transfection to the producer cells. In this system, the procedures were complicated and time-consuming, and approximately 30% of the vectors in the library were defective with no displaying peptide. These may hinder further extensive exploration of cancer-targeting vectors. To resolve these problems, in this study, we developed a novel method with the transfection of a fiber-modified plasmid library and a fiberless adenoviral DNA-TPC in Cre-expressing 293 cells. The use of in-cell Cre recombination and fiberless adenovirus greatly simplified the library-making steps. The fiberless adenovirus was useful in suppressing the expansion of unnecessary adenovirus vectors. In addition, the complexity of the library was more than a 10(4) level in one well in a 6-well dish, which was 10-fold higher than that of the original method. The results demonstrated that this novel method is useful in producing a high quality live adenovirus library, which could facilitate the development of targeted adenovirus vectors for a variety of applications in medicine.
authors.affiliation: Division of Gene and Immune Medicine, National Cancer Center Research Institute , 5-1-1 Tsukiji, Chuo-ku, Tokyo 104-0045, Japan.
authors.forename: Yuki
authors.initials: Y
authors.lastname: Yamamoto
authors.forename: Naoko
authors.initials: N
authors.lastname: Goto
authors.forename: Kazuki
authors.initials: K
authors.lastname: Miura
authors.forename: Kenta
authors.initials: K
authors.lastname: Narumi
authors.forename: Shumpei
authors.initials: S
authors.lastname: Ohnami
authors.forename: Hiroaki
authors.initials: H
authors.lastname: Uchida
authors.forename: Yoshiaki
authors.initials: Y
authors.lastname: Miura
authors.forename: Masato
authors.initials: M
authors.lastname: Yamamoto
authors.forename: Kazunori
authors.initials: K
authors.lastname: Aoki
mesh_terms: D000230:Adenocarcinoma / Q000235:genetics / Q000821:virology; D000256:Adenoviridae / Q000235:genetics*; D000257:Adenoviridae Infections / Q000235:genetics / Q000821:virology; D036022:Capsid Proteins / Q000235:genetics*; D004279:DNA, Viral / Q000235:genetics*; D005818:Genetic Engineering; D005822:Genetic Vectors / Q000235:genetics*; D016679:Genome, Viral; D057809:HEK293 Cells; D006801:Humans; D019426:Integrases / Q000378:metabolism; D008175:Lung Neoplasms / Q000235:genetics / Q000821:virology; D010446:Peptide Fragments / Q000378:metabolism*; D019151:Peptide Library; D010957:Plasmids / Q000008:administration & dosage; D014161:Transduction, Genetic
chemical_list: D036022:Capsid Proteins; D004279:DNA, Viral; D010446:Peptide Fragments; D019151:Peptide Library; C018315:hexon capsid protein, Adenovirus; C045073:Cre recombinase; D019426:Integrases
keywords: Cre recombination; adenovirus; fiberless; library; targeting vector