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Fowl adenovirus (FAdV)\u2010based vector might represent an alternative.<\/jats:p><\/jats:sec><jats:sec><jats:title>Methods<\/jats:title><jats:p>An intermediate plasmid containing FAdV\u20104 fiber genes, pMD\u2010FAV4Fs, was separated from FAdV\u20104 adenoviral plasmid pKFAV4GFP. An overlap extension polymerase chain reaction (PCR) was employed for fiber modification in pMD\u2010FAV4Fs, and the modified fibers were restored to generate new adenoviral plasmids through restriction\u2010assembly. FAdV\u20104 vectors were rescued and amplified in chicken LMH cells. Fluorescence microscopy and flow cytometry were used to evaluate the gene transfer efficiency. The amount of viruses binding to cells was determined by a real\u2010time PCR. A plaque\u2010forming assay and one\u2010step growth curve were used to evaluate virus growth.<\/jats:p><\/jats:sec><jats:sec><jats:title>Results<\/jats:title><jats:p>Four sites in the CD\u2010, DE\u2010, HI\u2010 and IJ\u2010loop of fiber1 knob could tolerate the insertion of exogenous peptide. The insertion of RGD4C peptide in the fiber1 knob significantly promoted FAdV\u20104 transduction to human adherent cells such as 293, A549 and HEp\u20102, and the insertion to the IJ\u2010loop demonstrated the best performance. The replacement of the fiber2 knob of FAdV\u20104 with that of HAdV\u201035 improved the gene transfer to human suspension cells such as Jurkat, K562 and U937. Fiber\u2010modified FAdV\u20104 vectors could transduce approximately 80% human cells at an acceptable multiplicity of infection. Enhanced gene transfer mainly resulted from increased virus binding. Fiber modifications did not significantly influence the growth of recombinant FAdV\u20104 in packaging cells.<\/jats:p><\/jats:sec><jats:sec><jats:title>Conclusions<\/jats:title><jats:p>As a proof of principle, it was feasible to enhance gene transduction of FAdV\u20104 vectors to human cells by modifying the fibers.<\/jats:p><\/jats:sec>","DOI":"10.1002\/jgm.3368","type":"journal-article","created":{"date-parts":[[2021,5,29]],"date-time":"2021-05-29T03:44:52Z","timestamp":1622259892000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":13,"title":["Fiber modifications enable fowl adenovirus 4 vectors to transduce human cells"],"prefix":"10.1002","volume":"23","author":[{"given":"Wenfeng","family":"Zhang","sequence":"first","affiliation":[{"name":"School of Laboratory Medicine Weifang Medical University  Weifang China"},{"name":"State Key Laboratory of Infectious Disease Prevention and Control National Institute for Viral 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