
The second parameter is the choice of targeting peptide. The site must be such that (i) assembly and packaging of the virus capsid are not affected and (ii) peptides must be exposed on the surface of the virus. The first decision is to choose the location for insertions. A combination of two important parameters will determine the success of genetically modified capsids for retargeting viral vectors in vivo. It has been shown that the accessibility of incorporated peptides can also be affected by the composition of flanking residues (J.

They also identified regions involved in heparin binding and were able to demonstrate altered tropism of viruses engineered to contain the serpin receptor ligand. reported a broad mutational analysis of the AAV-2 capsid that identified several sites amenable to the incorporation of peptides (35). Mutants fell into three pheno- typic groups, depending on their ability to assemble capsids, package DNA, and transduce cells. employed linker insertional mutagenesis to place small peptide sequences (three to five amino acids) randomly across the entire capsid coding region (34). In three mutants the peptide was exposed on the capsid surface and one of these showed preferential transduction of in- tegrin-expressing cells. A 14-amino-acid targeting peptide (L14) containing an RGD sequence was inserted into these sites and virus was produced. One study sug- gested six possible sites in the AAV capsid that were pre- dicted to be within surface loops (33). Several recent reports have begun to address structural features of AAV capsids in order to identify sites amenable to incorporation of peptides. The sequences of AAV serotypes 2, 1, 3, 4, and 5 are aligned, and regions modified in this study are shown enclosed by dashed boxes. (B) Alignments of putative Loops III and IV of the AAV serotypes. Numbering of mutations and sites match the corresponding papers. Sites of insertions are marked with an arrow (33), filled arrowhead (34), and open arrowhead (35). Published mutations and their corresponding sites are indicated and part of the  -barrel is shown with a horizontal arrow. Antigenic regions within the loops are enclosed by boxes that are numbered to the right (see text for details and references). Identical sequences (dark blue) and similar sequences (light blue) are highlighted. The alignments are shown for four domains (I–IV) within the capsid protein, which contain the antigenic loops of CPV.

#Make alignments macvector software#
494746, were aligned using the ClustalW alignment tool of the software program MacVector (Oxford Molecular Group, London). 494031 and canine parvovirus (CPV), GP No. 2982110 feline panleukopenia virus (FPV), GP No. 4092542 minute virus of mice (MVM), GP No. (A) The amino acid sequences of the capsid proteins of AAV-2, Genpept Accession No. Homology alignments of parvovirus Cap proteins.
