Light-Inducible Spatiotemporal Control of Gene Activation by Customizable Zinc Finger Transcription Factors

TLDR

Advanced gene regulatory systems are needed for research, synthetic biology, and gene‑based medicine, and an ideal system would allow tunable, reversible, repeatable, and spatially precise manipulation of gene expression across diverse DNA sequences. The authors engineered a light‑inducible transcription system that combines light‑sensitive proteins with programmable zinc finger transcription factors to meet these criteria. The LITEZ system employs two Arabidopsis thaliana light‑inducible dimerizers, GIGANTEA and the LOV domain of FKF1, to control engineered zinc finger transcription factor activity in human cells. In human cells, LITEZ enables reversible and repeatable gene activation by adjusting illumination duration, intensity‑dependent expression levels, and spatially defined patterns via photomasks, thereby providing precise regulation for biotechnology and medical applications.

Abstract

Advanced gene regulatory systems are necessary for scientific research, synthetic biology, and gene-based medicine. An ideal system would allow facile spatiotemporal manipulation of gene expression within a cell population that is tunable, reversible, repeatable, and can be targeted to diverse DNA sequences. To meet these criteria, a gene regulation system was engineered that combines light-sensitive proteins and programmable zinc finger transcription factors. This system, light-inducible transcription using engineered zinc finger proteins (LITEZ), uses two light-inducible dimerizing proteins from Arabidopsis thaliana, GIGANTEA and the LOV domain of FKF1, to control synthetic zinc finger transcription factor activity in human cells. Activation of gene expression in human cells engineered with LITEZ was reversible and repeatable by modulating the duration of illumination. The level of gene expression could also be controlled by modulating light intensity. Finally, gene expression could be activated in a spatially defined pattern by illuminating the human cell culture through a photomask of arbitrary geometry. LITEZ enables new approaches for precisely regulating gene expression in biotechnology and medicine, as well as studying gene function, cell-cell interactions, and tissue morphogenesis.