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Optogenetic Inhibitor of the Transcription Factor CREB

Ahmed M Ali1, Jakeb M Reis2, Yan Xia3

  • 1Department of Chemistry, University of Toronto, 80 St. George Street, Toronto, ON M5S 3H6, Canada; Department of Medicinal Chemistry, Faculty of Pharmacy, Assiut University, Assiut 71515, Egypt.

Chemistry & Biology
|November 23, 2015
PubMed

Insights

This study introduces opto-DN-CREB, a novel optogenetic tool for controlling gene expression. Blue light activates CREB (cAMP response element-binding protein) by modulating a dominant-negative inhibitor, enabling precise regulation of transcription.

Area of Science:

  • Molecular Biology
  • Neuroscience
  • Optogenetics

Background:

  • Current optogenetic transcription control lacks mimicry of endogenous factors.
  • Endogenous transcription factors exhibit complex, multi-site genomic interactions.
  • Dominant-negative inhibitors offer a pathway to mimic natural gene regulation.

Purpose of the Study:

  • To develop a novel optogenetic system for precise transcription factor control.
  • To create a blue-light-inducible dominant-negative inhibitor for CREB.
  • To demonstrate spatiotemporal regulation of CREB activity and its downstream targets.

Main Methods:

  • Fusion of A-CREB (dominant-negative CREB inhibitor) with photoactive yellow protein (PYP).
  • Utilizing PYP's light-induced conformational change to regulate A-CREB/CREB interaction.
  • In vitro, cell-based (HEK293T), and neuronal experiments to validate functionality.

Main Results:

  • Opto-DN-CREB demonstrated blue-light-dependent inhibition of CREB activity.
  • Activation of CREB was achieved by preventing A-CREB/CREB coiled-coil formation.
  • Controlled expression of CREB target genes (NR4A2, c-Fos) in neurons via blue light.

Conclusions:

  • Opto-DN-CREB provides a generalizable method for optogenetic control of transcription factors.
  • This approach mimics natural gene regulation by controlling dominant-negative inhibitors.
  • Offers a versatile platform for spatiotemporal manipulation of native transcriptional events.
Keywords:
CREBPYPdominant negativeendogenousgenetically encodedoptogeneticsphoto-controlphotoactive yellow proteinphotoisomerizationtranscription factor

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