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@ARTICLE{Uenaka:285483,
      author       = {Uenaka, Takeshi and Napole, Alan and Saha, Aninda Dibya and
                      Sun, Duo and Singavarapu, Angelina and Calzada, Elizabeth
                      and Chen, Jiahui and Erlebach, Lena and McQuade, Amanda and
                      Ramos, Daniel M and Rigamonti, Alessandra and Salazar, Lisa
                      and Samelson, Avi J and Sedov, Kamilla and Welsh, Natalie J
                      and Wild, Katleen and Wu, Qianxin and Arenas, Ernest and
                      Bassett, Andrew R and Kampmann, Martin and
                      Kronenberg-Versteeg, Deborah and Merkle, Florian T and
                      Schüle, Birgitt and Thompson, Leslie M and Skarnes, William
                      C and Ward, Michael E and Wernig, Marius},
      title        = {{P}revention of transgene silencing during human
                      pluripotent stem cell differentiation.},
      journal      = {Cell stem cell},
      volume       = {33},
      number       = {3},
      issn         = {1934-5909},
      address      = {Amsterdam [u.a.]},
      publisher    = {Elsevier},
      reportid     = {DZNE-2026-00259},
      pages        = {517 - 530.e8},
      year         = {2026},
      abstract     = {Transgenes are often silenced upon differentiation of
                      pluripotent stem cells using conventional expression
                      systems. Here, we developed the TK4 PiggyBac vector to
                      conduct a comparative analysis to evaluate the impact of
                      various promoters, transcriptional regulatory elements,
                      insulators, and genomic integration sites on transgene
                      silencing during neuronal differentiation. Our findings
                      reveal that specific combinations of CAG and Ubc promoters
                      with the Woodchuck hepatitis virus post-transcriptional
                      regulatory element (WPRE) can prevent transgene silencing
                      during differentiation, whereas chromatin insulators have
                      less impact on sustained expression. Three novel safe harbor
                      loci, distant from known genes, as well as the citrate lyase
                      beta-like (CLYBL) locus, similarly support the prevention of
                      transgene silencing. Remarkably, the TK4 vector showed
                      complete resistance to silencing across various neuronal and
                      microglial differentiation protocols, as independently
                      confirmed by seven laboratories. This construct will be
                      highly useful for assays requiring stable transgene
                      expression during differentiation and holds potential for
                      broad applications in various research fields.},
      keywords     = {Humans / Cell Differentiation: genetics / Transgenes:
                      genetics / Pluripotent Stem Cells: cytology / Pluripotent
                      Stem Cells: metabolism / Gene Silencing / Promoter Regions,
                      Genetic: genetics / Genetic Vectors: genetics / Neurons:
                      cytology / Neurons: metabolism / PiggyBac vector (Other) /
                      UCOE (Other) / WPRE (Other) / chromatin insulator (Other) /
                      human pluripotent stem cells (Other) / microglia (Other) /
                      neuron (Other) / promoter activity (Other) / safe harbor
                      locus (Other) / transgene silencing (Other) / ubiquitous
                      chromatin opening element (Other) / woodchuck hepatitis
                      posttranscriptional regulatory element (Other)},
      cin          = {AG Jucker / AG Neher (Tübingen) / AG Kronenberg-Versteeg},
      ddc          = {570},
      cid          = {I:(DE-2719)1210001 / I:(DE-2719)1210012 /
                      I:(DE-2719)1210015},
      pnm          = {352 - Disease Mechanisms (POF4-352) / 351 - Brain Function
                      (POF4-351)},
      pid          = {G:(DE-HGF)POF4-352 / G:(DE-HGF)POF4-351},
      typ          = {PUB:(DE-HGF)16},
      pubmed       = {pmid:41690310},
      doi          = {10.1016/j.stem.2026.01.007},
      url          = {https://pub.dzne.de/record/285483},
}