Prolonged activity of the transposase helper may raise safety concerns during DNA transposon-based gene therapy
DNA transposon-based gene delivery vectors represent a promising new branch of randomly integrating vector development for gene therapy. For the side-by-side evaluation of the piggyBac and Sleeping Beauty systems—the only DNA transposons currently employed in clinical trials—during therapeutic intervention, we treated the mouse model of tyrosinemia type I with liver-targeted gene delivery using both transposon vectors. For genome-wide mapping of transposon insertion sites we developed a new next-generation sequencing procedure called streptavidin-based enrichment sequencing, which allowed us to identify approximately one million integration sites for both systems. We revealed that a high proportion of piggyBac integrations are clustered in hot regions and found that they are frequently recurring at the same genomic positions among treated animals, indicating that the genome-wide distribution of Sleeping Beauty-generated integrations is closer to random. We also revealed that the piggyBac transposase protein exhibits prolonged activity, which predicts the risk of oncogenesis by generating chromosomal double-strand breaks. Safety concerns associated with prolonged transpositional activity draw attention to the importance of squeezing the active state of the transposase enzymes into a narrower time window.
Molecular Therapy - Methods and Clinical Development
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Imre, Gergely; Takács, Bertalan; Czipa, Erik; Drubi, Andrea Bakné; Jaksa, Gábor; Latinovics, Dóra; Nagy, Andrea; Karkas, Réka; Hudoba, Liza; Vásárhelyi, Bálint Márk; Pankotai-Bodó, Gabriella; Blastyák, András; Hegedűs, Zoltán; Germán, Péter; Bálint, Balázs; Ahmed Abdullah, Khaldoon Sadiq; Kopasz, Anna Georgina; Kovács, Anita; Nagy, László G.; Sükösd, Farkas; Pintér, Lajos; Rülicke, Thomas; Barta, Endre; Nagy, István; Haracska, Lajos; and Mátés, Lajos, "Prolonged activity of the transposase helper may raise safety concerns during DNA transposon-based gene therapy" (2023). Kean Publications. 122.