Skip to content
DraconiticResearch
Search Sources API
189,280 studies31 sourcesUpdated 50 min ago
PreprintCC BY-NCbioRxiv

Genetic control couples TGFβ-independent pluripotent stem cell renewal to deterministic forward programming across germ layers

  • S. Bottini
  • F. Amodeo
  • D. Avnoor
  • M. De Gregorio
  • M. Arigoni
  • F. Bachinger
  • P. Barrett
  • D. L. Mack
  • P. E. Porporato
  • P. Defilippi
  • L. Vallier
  • A. S. Smith
2 more authors
  • E. Balmas
  • A. Bertero1
  1. 1University of Turin
Read full textPDFDOICite

Abstract

Reproducible and scalable manufacture of human induced pluripotent stem cell (hiPSC) derivatives requires control over expansion and differentiation. Transcription factor-mediated forward programming can place hiPSC differentiation under genetic control, but expansion still relies on extracellular growth-factor supplementation, increasing process complexity and operator dependence. Here we introduce GERALT (Genetically Enhanced Renewal and Autonomous Lineage Transdifferentiation), a push-pull genetic circuit assembled through targeted safe-harbour integration that couples NANOG-supported, TGF{beta}-independent renewal to doxycycline-inducible lineage programming and simultaneous NANOG silencing. GERALT maintained pluripotency during extended TGF{beta} withdrawal in both adherent and suspension culture, yet remained programmable towards myogenic, neuronal, and hepatic fates. Compared with the established OPTi-OX platform in myogenesis, GERALT enabled differentiation in simplified medium while canalising and accelerating commitment. GERALT also supported direct myogenic programming in suspension and enabled direct casting of hiPSCs into hydrogel followed by in-tissue programming, yielding electrically responsive engineered muscle tissues. These results extend genetic control upstream into stem cell renewal, facilitating deployment of new approach methodologies and other biomanufacturing applications.

Related

  1. Impacts of different SETD1A mutations and genetic backgrounds on human induced pluripotent stem cell-derived hippocampal neurons
    Y. Hong, T. Zhao, Q. Yang +15
    bioRxivPreprintCC BY-NC-NDFull textNeurosciencebioRxiv
  2. DiffDose: Differentiable Programming for Personalized Dose-Regimen Optimal Control
    S. Hajhashemi, A. Emad, M. Craig
    bioRxivPreprintCC BY-NCFull textSystems & synthetic biologybioRxiv
  3. Inducible cAMP Early Repressor Limits Glycolytic Programming in Regulatory T Cells and Supports Control of Inflammatory Colitis.
    Aleksandra Lopez Krol, Luana Silva, Daniele C Nascimento +10
    European journal of immunologyPeer-reviewedCC BYFull textImmunologyEurope PMC
  4. A twin delayed deep deterministic-based control method for a full vehicle semi-active suspension system
    Yongjun Wan, Xiaoming Wang, Gang Zhi +4
    PLOS OnePeer-reviewedCC BY 4.0Full textNeurosciencePLOS
  5. Behavioral adaptation to noise is independent of age and cognitive inhibitory control
    Sónia L. Coelho-de-Sousa, Miriam I. Marrufo-Pérez, Enrique A. Lopez-Poveda
    Scientific ReportsPeer-reviewedCC BYFull textNeuroscienceDOAJ

Cite

BibTeXRIS
BibTeX
@misc{bottini2026genetic,
  title = {{Genetic control couples TGFβ-independent pluripotent stem cell renewal to deterministic forward programming across germ layers}},
  author = {S. Bottini and F. Amodeo and D. Avnoor and M. De Gregorio and M. Arigoni and F. Bachinger and P. Barrett and D. L. Mack and P. E. Porporato and P. Defilippi and L. Vallier and A. S. Smith and E. Balmas and A. Bertero},
  howpublished = {bioRxiv},
  year = {2026},
  month = oct,
  doi = {10.64898/2026.10.25.761636},
  url = {https://www.biorxiv.org/content/10.64898/2026.10.25.761636v1},
  note = {CC BY-NC},
}
RIS
TY  - UNPB
TI  - Genetic control couples TGFβ-independent pluripotent stem cell renewal to deterministic forward programming across germ layers
AU  - S. Bottini
AU  - F. Amodeo
AU  - D. Avnoor
AU  - M. De Gregorio
AU  - M. Arigoni
AU  - F. Bachinger
AU  - P. Barrett
AU  - D. L. Mack
AU  - P. E. Porporato
AU  - P. Defilippi
AU  - L. Vallier
AU  - A. S. Smith
AU  - E. Balmas
AU  - A. Bertero
PY  - 2026
DA  - 2026/10/25
PB  - bioRxiv
DO  - 10.64898/2026.10.25.761636
UR  - https://www.biorxiv.org/content/10.64898/2026.10.25.761636v1
AB  - Reproducible and scalable manufacture of human induced pluripotent stem cell (hiPSC) derivatives requires control over expansion and differentiation. Transcription factor-mediated forward programming can place hiPSC differentiation under genetic control, but expansion still relies on extracellular growth-factor supplementation, increasing process complexity and operator dependence. Here we introduce GERALT (Genetically Enhanced Renewal and Autonomous Lineage Transdifferentiation), a push-pull genetic circuit assembled through targeted safe-harbour integration that couples NANOG-supported, TGF{beta}-independent renewal to doxycycline-inducible lineage programming and simultaneous NANOG silencing. GERALT maintained pluripotency during extended TGF{beta} withdrawal in both adherent and suspension culture, yet remained programmable towards myogenic, neuronal, and hepatic fates. Compared with the established OPTi-OX platform in myogenesis, GERALT enabled differentiation in simplified medium while canalising and accelerating commitment. GERALT also supported direct myogenic programming in suspension and enabled direct casting of hiPSCs into hydrogel followed by in-tissue programming, yielding electrically responsive engineered muscle tissues. These results extend genetic control upstream into stem cell renewal, facilitating deployment of new approach methodologies and other biomanufacturing applications.
ER  -