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Light-inducible gene regulation with engineered zinc finger proteins.

Publication ,  Journal Article
Polstein, LR; Gersbach, CA
Published in: Methods in molecular biology (Clifton, N.J.)
January 2014

The coupling of light-inducible protein-protein interactions with gene regulation systems has enabled the control of gene expression with light. In particular, heterodimer protein pairs from plants can be used to engineer a gene regulation system in mammalian cells that is reversible, repeatable, tunable, controllable in a spatiotemporal manner, and targetable to any DNA sequence. This system, Light-Inducible Transcription using Engineered Zinc finger proteins (LITEZ), is based on the blue light-induced interaction of GIGANTEA and the LOV domain of FKF1 that drives the localization of a transcriptional activator to the DNA-binding site of a highly customizable engineered zinc finger protein. This chapter provides methods for modifying LITEZ to target new DNA sequences, engineering a programmable LED array to illuminate cell cultures, and using the modified LITEZ system to achieve spatiotemporal control of transgene expression in mammalian cells.

Duke Scholars

Published In

Methods in molecular biology (Clifton, N.J.)

DOI

EISSN

1940-6029

ISSN

1064-3745

Publication Date

January 2014

Volume

1148

Start / End Page

89 / 107

Related Subject Headings

  • Zinc Fingers
  • Transcriptional Activation
  • Transcription, Genetic
  • Transcription Factors
  • Protein Engineering
  • Promoter Regions, Genetic
  • Photochemical Processes
  • Light
  • Humans
  • Hela Cells
 

Citation

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Polstein, L. R., & Gersbach, C. A. (2014). Light-inducible gene regulation with engineered zinc finger proteins. Methods in Molecular Biology (Clifton, N.J.), 1148, 89–107. https://doi.org/10.1007/978-1-4939-0470-9_7
Polstein, Lauren R., and Charles A. Gersbach. “Light-inducible gene regulation with engineered zinc finger proteins.Methods in Molecular Biology (Clifton, N.J.) 1148 (January 2014): 89–107. https://doi.org/10.1007/978-1-4939-0470-9_7.
Polstein LR, Gersbach CA. Light-inducible gene regulation with engineered zinc finger proteins. Methods in molecular biology (Clifton, NJ). 2014 Jan;1148:89–107.
Polstein, Lauren R., and Charles A. Gersbach. “Light-inducible gene regulation with engineered zinc finger proteins.Methods in Molecular Biology (Clifton, N.J.), vol. 1148, Jan. 2014, pp. 89–107. Epmc, doi:10.1007/978-1-4939-0470-9_7.
Polstein LR, Gersbach CA. Light-inducible gene regulation with engineered zinc finger proteins. Methods in molecular biology (Clifton, NJ). 2014 Jan;1148:89–107.

Published In

Methods in molecular biology (Clifton, N.J.)

DOI

EISSN

1940-6029

ISSN

1064-3745

Publication Date

January 2014

Volume

1148

Start / End Page

89 / 107

Related Subject Headings

  • Zinc Fingers
  • Transcriptional Activation
  • Transcription, Genetic
  • Transcription Factors
  • Protein Engineering
  • Promoter Regions, Genetic
  • Photochemical Processes
  • Light
  • Humans
  • Hela Cells