Key result
A single AD domain of the inverted terminal repeat serves as a minimal origin of circular adeno-associated virus replication and is an efficient template for encapsidation of single-stranded DNA genomes.
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The identification of the AD domain as a minimal origin for circular AAV replication provides new insights into AAV replication and packaging mechanisms.
A novel pathway of adeno-associated virus (AAV) replication marked by the assembly of circular monomer duplex intermediates (cAAV) has been recently discovered. In the present report we identify a single AD domain of the inverted terminal repeat as a minimal origin of cAAV replication. A small internal palindrome (BB'), necessary for optimal Rep-inverted terminal repeat interaction, does not contribute to the efficiency of cAAV replication, while the terminal resolution site is an essential cis-acting element. Furthermore, recombinant cAAV vectors that encompass only the AD domain replicate exclusively in a circular form and no detectable linear duplex replicative intermediates are generated, suggesting that both pathways of AAV replication are independent and can be separated. In addition, we show that cAAVs are efficient templates for encapsidation of single-stranded DNA genomes, an observation that assigns a biological role for these novel replication species. Together, these findings shed new light on the current model of AAV replication and packaging.
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Musatov et al. (2002) studied Adeno-associated virus (AAV) replication. AD domain of the inverted terminal repeat vs. Intact TRT domain or other ITR deletions was evaluated on cAAV replication and packaging. A single AD domain of the inverted terminal repeat serves as a minimal origin of circular adeno-associated virus replication and is an efficient template for encapsidation of single-stranded DNA genomes.
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