%0 Journal Article
%T Sleeping Beauty mRNA-LNP enables stable rAAV transgene expression in mouse and NHP hepatocytes and improves vector potency.
%A Zakas PM
%A Cunningham SC
%A Doherty A
%A van Dijk EB
%A Ibraheim R
%A Yu S
%A Mekonnen BD
%A Lang B
%A English EJ
%A Sun G
%A Duncan MC
%A Benczkowski MS
%A Altshuler RC
%A Singh MJ
%A Kibbler ES
%A Tonga GY
%A Wang ZJ
%A Wang ZJ
%A Li G
%A An D
%A Rottman JB
%A Bhavsar Y
%A Purcell C
%A Jain R
%A Alberry R
%A Roquet N
%A Fu Y
%A Citorik RJ
%A Rubens JR
%A Holmes MC
%A Cotta-Ramusino C
%A Querbes W
%A Alexander IE
%A Salomon WE
%J Mol Ther
%V 0
%N 0
%D 2024 Jul 2
%M 38981468
%F 12.91
%R 10.1016/j.ymthe.2024.06.021
%X Recombinant adeno-associated virus (rAAV) vector gene delivery systems have demonstrated great promise in clinical trials but continue to face durability and dose-related challenges. Unlike rAAV gene therapy, integrating gene addition approaches can provide curative expression in mitotically active cells and pediatric populations. We explored a novel in vivo delivery approach based on an engineered transposase, Sleeping Beauty (SB100X), delivered as an mRNA within a lipid nanoparticle (LNP), in combination with an rAAV-delivered transposable transgene. This combinatorial approach achieved correction of ornithine transcarbamylase deficiency in the neonatal Spfash mouse model following a single delivery to dividing hepatocytes in the newborn liver. Correction remained stable into adulthood, while a conventional rAAV approach resulted in a return to the disease state. In non-human primates, integration by transposition, mediated by this technology, improved gene expression 10-fold over conventional rAAV-mediated gene transfer while requiring 5-fold less vector. Additionally, integration site analysis confirmed a random profile while specifically targeting TA dinucleotides across the genome. Together, these findings demonstrate that transposable elements can improve rAAV-delivered therapies by lowering the vector dose requirement and associated toxicity while expanding target cell types.