Prolonged activity of the transposase helper may raise safety concerns during DNA transposon-based gene therapy.

Gergely Imre, Bertalan Takács, Erik Czipa, Andrea Bakné Drubi, Gábor Jaksa, Dóra Latinovics, Andrea Nagy, Réka Karkas, Liza Hudoba, Bálint Márk Vásárhelyi, Gabriella Pankotai-Bodó, András Blastyák, Zoltán Hegedűs, Péter Germán, Balázs Bálint, Khaldoon Sadiq Ahmed Abdullah, Anna Georgina Kopasz, Anita Kovács, László G Nagy, Farkas Sükösd, Lajos Pintér, Thomas Rülicke, Endre Barta, István Nagy, Lajos Haracska, Lajos Mátés

Molecular therapy. Methods & clinical development 2023 Jun 08

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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. © 2023 The Author(s).

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PMID: 37025950

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