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Bidirectional approaches for optogenetic regulation of gene expression in mammalian cells using Arabidopsis cryptochrome 2.

Publication ,  Journal Article
Pathak, GP; Spiltoir, JI; Höglund, C; Polstein, LR; Heine-Koskinen, S; Gersbach, CA; Rossi, J; Tucker, CL
Published in: Nucleic acids research
November 2017

Optogenetic tools allow regulation of cellular processes with light, which can be delivered with spatiotemporal resolution. In previous work, we used cryptochrome 2 (CRY2) and CIB1, Arabidopsis proteins that interact upon light illumination, to regulate transcription with light in yeast. While adopting this approach to regulate transcription in mammalian cells, we observed light-dependent redistribution and clearing of CRY2-tethered proteins within the nucleus. The nuclear clearing phenotype was dependent on the presence of a dimerization domain contained within the CRY2-fused transcriptional activators. We used this knowledge to develop two different approaches to regulate cellular protein levels with light: a system using CRY2 and CIB1 to induce protein expression with light through stimulation of transcription, and a system using CRY2 and a LOV-fused degron to simultaneously block transcription and deplete protein levels with light. These tools will allow precise, bi-directional control of gene expression in a variety of cells and model systems.

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Published In

Nucleic acids research

DOI

EISSN

1362-4962

ISSN

0305-1048

Publication Date

November 2017

Volume

45

Issue

20

Start / End Page

e167

Related Subject Headings

  • Zebrafish
  • Transcriptional Activation
  • Optogenetics
  • Light
  • Humans
  • HEK293 Cells
  • Gene Expression Regulation
  • Developmental Biology
  • DNA-Binding Proteins
  • Cryptochromes
 

Citation

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ICMJE
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Pathak, G. P., Spiltoir, J. I., Höglund, C., Polstein, L. R., Heine-Koskinen, S., Gersbach, C. A., … Tucker, C. L. (2017). Bidirectional approaches for optogenetic regulation of gene expression in mammalian cells using Arabidopsis cryptochrome 2. Nucleic Acids Research, 45(20), e167. https://doi.org/10.1093/nar/gkx260
Pathak, Gopal P., Jessica I. Spiltoir, Camilla Höglund, Lauren R. Polstein, Sari Heine-Koskinen, Charles A. Gersbach, Jari Rossi, and Chandra L. Tucker. “Bidirectional approaches for optogenetic regulation of gene expression in mammalian cells using Arabidopsis cryptochrome 2.Nucleic Acids Research 45, no. 20 (November 2017): e167. https://doi.org/10.1093/nar/gkx260.
Pathak GP, Spiltoir JI, Höglund C, Polstein LR, Heine-Koskinen S, Gersbach CA, et al. Bidirectional approaches for optogenetic regulation of gene expression in mammalian cells using Arabidopsis cryptochrome 2. Nucleic acids research. 2017 Nov;45(20):e167.
Pathak, Gopal P., et al. “Bidirectional approaches for optogenetic regulation of gene expression in mammalian cells using Arabidopsis cryptochrome 2.Nucleic Acids Research, vol. 45, no. 20, Nov. 2017, p. e167. Epmc, doi:10.1093/nar/gkx260.
Pathak GP, Spiltoir JI, Höglund C, Polstein LR, Heine-Koskinen S, Gersbach CA, Rossi J, Tucker CL. Bidirectional approaches for optogenetic regulation of gene expression in mammalian cells using Arabidopsis cryptochrome 2. Nucleic acids research. 2017 Nov;45(20):e167.
Journal cover image

Published In

Nucleic acids research

DOI

EISSN

1362-4962

ISSN

0305-1048

Publication Date

November 2017

Volume

45

Issue

20

Start / End Page

e167

Related Subject Headings

  • Zebrafish
  • Transcriptional Activation
  • Optogenetics
  • Light
  • Humans
  • HEK293 Cells
  • Gene Expression Regulation
  • Developmental Biology
  • DNA-Binding Proteins
  • Cryptochromes