Drug Delivery
RECOMBINANT AAV CONSTRUCTS FOR INCREASED TRANSGENE EXPRESSION
WARF: P230309US02
Inventors: Kinjal Majumder, Rhiannon Abrahams
The Invention
UW-Madison researchers have discovered rearrangement sequences that can be used to engineer rAAV vectors so that they may assemble larger therapeutic transgenes. Using forward and reverse primers that are oriented away from the main AAV genome, the inventors  have identified the recombination junction between two successive AAV genomes that have recombined in a head-to-tail orientation, amplified the putative recombined junction, cloned it into plasmids via TA-cloning and Sanger sequenced the recombination junction. This has enabled the inventors to characterize the exact sequence that is formed between two linearly recombined AAV genomes. The inventors’ findings both characterize the DNA junction and implicate the molecular machinery (i.e., the DNA repair protein PARP1) that regulates AAV rearrangement. Current and future studies are aimed at high-throughput sequencing the entire spectrum of AAV recombinants and assessing the rearrangement efficiency in mutant AAV viruses, including assessing whether the substantially modified rAAV vectors (which are stripped to essentials) include or can be engineered with sequences to facilitate or direct linear recombination. Understanding how to engineer rAAV vectors to facilitate the assembly of rAAV sequences could support the production of a much larger gene product than can currently be efficiently produced.
For current licensing status, please contact Jennifer Gottwald at [javascript protected email address] or 608-960-9854