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US 12,600,984 B2Granted
Viral and non-viral nanoplasmid vectors with improved production
AssigneeAldevron, LLC
InventorJames A. Williams
Life of this patent
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AssigneeAldevron, LLC
InventorJames A. Williams
Estimating the term…
A method for improving the replication of a covalently closed circular plasmid is provided. The method includes providing a covalently closed circular plasmid having a Pol I-dependent origin of replication, and an insert including a structured DNA sequence selected from inverted repeat sequences, direct repeat sequences, homopolymeric repeat sequences, eukaryotic origins of replication or eukaryotic promoter enhancer sequences, wherein the structured DNA sequence is located at a distance of less than 1000 bp from the Pol I-dependent origin of replication in the direction of replication. The method also includes modifying the covalently closed circular recombinant molecule such that the Pol I-dependent origin of replication is replaced with a Pol III-dependent origin of replication, whereby the resultant Pol III-dependent origin of replication covalently closed circular plasmid has improved replication. An antibiotic marker free covalently closed circular recombinant DNA molecule is also provided.
A method for replicating a covalently closed circular plasmid comprising the following steps:
A covalently closed circular plasmid comprising a backbone and an insert, the backbone comprising a Pol III-dependent R6K origin of replication and an RNA-OUT RNA selectable marker, wherein the backbone is less than 1000 bp, and the insert comprises a structured DNA sequence selected from the group consisting of an inverted repeat sequence, a direct repeat sequence, and a eukaryotic origin of replication, wherein the structured DNA sequence is located at a distance of less than 1000 bp from the Pol III-dependent R6K origin of replication in the direction of replication.
Dependencies are inferred from the claim text.