Plug-and-Play or Plug-and-Pray? Impact of random versus targeted integration on the linearity and orthogonality of synthetic gene circuits in CHO cells.
- 1School of Chemical & Bioprocess Engineering, University College Dublin, D04 V1W8, Ireland.
- 2Biotechnology Research Institute for the Green Transition (BRIGHT), Technical University of Denmark, Kgs. Lyngby, 2800, Denmark.
- 3Australian Institute for Bioengineering and Nanotechnology, University of Queensland, Brisbane, 4072, Australia.
Abstract
Predictable and tuneable gene expression is essential for advanced mammalian cell engineering yet remains challenged by genomic position effects and copy number variability. This study systematically compares random integration (RI) and targeted integration (TI) of two synthetic lineariser gene circuits, TetR/eGFP_Lin and PhlFd/mCherry_Lin, in CHO-S cells. Both circuits exhibited strong linearity and orthogonality across integration strategies, but integration method and repressor choice produced distinct performance trade-offs. RI increased achievable dynamic range for PhlFd but increased basal leakiness for TetR, whereas TI, via a dual-landing-pad system, constrained copy number, and reduced basal expression and PhlFd expression variability. These findings highlight integration context and circuit architecture as key design parameters informing rational design of synthetic regulators for biopharmaceutical applications.
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BibTeX
@article{lim2026plugandplay,
title = {{Plug-and-Play or Plug-and-Pray? Impact of random versus targeted integration on the linearity and orthogonality of synthetic gene circuits in CHO cells.}},
author = {Sheryl Li Yan Lim and Lars K Nielsen and Ioscani Jiménez Del Val and Jesús Lavado-García},
journal = {Synthetic and systems biotechnology},
year = {2026},
month = oct,
doi = {10.1016/j.synbio.2026.10.004},
url = {https://europepmc.org/article/MED/42926929},
note = {CC BY-NC-ND},
}
RIS
TY - JOUR
TI - Plug-and-Play or Plug-and-Pray? Impact of random versus targeted integration on the linearity and orthogonality of synthetic gene circuits in CHO cells.
AU - Sheryl Li Yan Lim
AU - Lars K Nielsen
AU - Ioscani Jiménez Del Val
AU - Jesús Lavado-García
PY - 2026
DA - 2026/10/15
JO - Synthetic and systems biotechnology
DO - 10.1016/j.synbio.2026.10.004
UR - https://europepmc.org/article/MED/42926929
AB - Predictable and tuneable gene expression is essential for advanced mammalian cell engineering yet remains challenged by genomic position effects and copy number variability. This study systematically compares random integration (RI) and targeted integration (TI) of two synthetic lineariser gene circuits, TetR/eGFP_Lin and PhlFd/mCherry_Lin, in CHO-S cells. Both circuits exhibited strong linearity and orthogonality across integration strategies, but integration method and repressor choice produced distinct performance trade-offs. RI increased achievable dynamic range for PhlFd but increased basal leakiness for TetR, whereas TI, via a dual-landing-pad system, constrained copy number, and reduced basal expression and PhlFd expression variability. These findings highlight integration context and circuit architecture as key design parameters informing rational design of synthetic regulators for biopharmaceutical applications.
SN - 2405-805X
ER -