title: Sites in the AAV5 capsid tolerant to deletions and tandem duplications.
abstract: Gene therapy vectors based on adeno-associated virus (AAV) have shown much promise in clinical trials for the treatment of a variety of diseases. However, the ability to manipulate and engineer the viral surface for enhanced efficiency is necessary to overcome such barriers as pre-existing immunity and transduction of non-target cells that currently limit AAV applications. Although single amino acid changes and peptide insertions at select sites have been explored previously, the tolerance of AAV to small deletions and tandem duplications of sequence has not been globally addressed. Here, we have generated a large, diverse library of >10(5) members containing deletions and tandem duplications throughout the viral capsid of AAV5. Four unique mutants were identified that maintain the ability to form viral particles, with one showing improved transduction on both 293T and BEAS-2B cells. This approach may find potential use for the generation of novel variants with improved and altered properties or in the identification of sites that are tolerant to insertions of targeting ligands.
authors.affiliation: Department of Biomedical Engineering, The Johns Hopkins University, Baltimore, MD 21218, USA.
authors.forename: Kaoru
authors.initials: K
authors.lastname: Hida
authors.forename: Sang Y
authors.initials: SY
authors.lastname: Won
authors.forename: Giovanni
authors.initials: G
authors.lastname: Di Pasquale
authors.forename: Justin
authors.initials: J
authors.lastname: Hanes
authors.forename: John A
authors.initials: JA
authors.lastname: Chiorini
authors.forename: Marc
authors.initials: M
authors.lastname: Ostermeier
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002213:Capsid / Q000737:chemistry / Q000378:metabolism*; D036022:Capsid Proteins / Q000737:chemistry / Q000378:metabolism; D002460:Cell Line; D000229:Dependovirus / Q000235:genetics* / Q000378:metabolism*; D015723:Gene Library; D005818:Genetic Engineering; D008958:Models, Molecular; D008969:Molecular Sequence Data; D010455:Peptides / Q000378:metabolism; D055503:Protein Multimerization; D020836:Protein Structure, Quaternary; D017384:Sequence Deletion; D020080:Tandem Repeat Sequences / Q000235:genetics*; D014161:Transduction, Genetic
chemical_list: D036022:Capsid Proteins; D010455:Peptides