Imre, Gergely (University of Szeged / Biological Research Centre Szeged) Gergely Imre Haracska, Lajos (Biological Research Centre Szeged / Delta Bio 2000 Ltd.) Lajos Haracska Kopasz, Anna Georgina (Biological Research Centre Szeged) Anna Georgina Kopasz Ahmed Abdullah, Khaldoon Sadiq (University of Szeged / Biological Research Centre Szeged) Khaldoon Sadiq Ahmed Abdullah Bálint, Balázs (Biological Research Centre Szeged) Balázs Bálint Germán, Péter (Biological Research Centre Szeged) Péter Germán Hegedűs, Zoltán (Biological Research Centre Szeged) Zoltán Hegedűs Blastyák, András (Biological Research Centre Szeged) András Blastyák Pankotai-Bodó, Gabriella (University of Szeged) Gabriella Pankotai-Bodó Vásárhelyi, Bálint Márk (Biological Research Centre Szeged) Bálint Márk Vásárhelyi Hudoba, Liza (Biological Research Centre Szeged) Liza Hudoba Hudoba, Liza (Biological Research Centre Szeged) Liza Hudoba Mátés, Lajos (Biological Research Centre Szeged) Lajos Mátés Karkas, Réka (University of Szeged / Biological Research Centre Szeged) Réka Karkas Nagy, Andrea (Biological Research Centre Szeged) Andrea Nagy Latinovics, Dóra (Seqomics Biotechnology Ltd) Dóra Latinovics Jaksa, Gábor (Delta Bio 2000 Ltd.) Gábor Jaksa Drubi, Andrea Bakné (University of Szeged / Biological Research Centre Szeged) Andrea Bakné Drubi Takács, Bertalan (Biological Research Centre Szeged) Bertalan Takács Czipa, Erik (University of Debrecen) Erik Czipa Nagy, István (Biological Research Centre Szeged) István Nagy Barta, Endre (University of Debrecen) Endre Barta Rülicke, Thomas (University of Veterinary Medicine Vienna) Thomas Rülicke Pintér, Lajos (Delta Bio 2000 Ltd.) Lajos Pintér Sükösd, Farkas (University of Szeged) Farkas Sükösd Nagy, László G. (Biological Research Centre Szeged) László G. Nagy Kovács, Anita (Wenzhou-Kean University / Chinese Academy of Sciences) Anita Kovács Cell Press 2023 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. PDFDocument en 2023-10-17T08:12:52.957Z Tyrosinemia Type-I; Sleeping-Beauty Transposase; Piggybac Transposon; Murine Model; Hepatic-Dysfunction; Site Selection; Mouse-Liver; Methylations; Mutagenesis; Expression 1977190 b application/pdf CC BY 4.0 International