Light-inducible gene regulation with engineered zinc finger proteins.
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
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Related Subject Headings
- Zinc Fingers
- Transcriptional Activation
- Transcription, Genetic
- Transcription Factors
- Protein Engineering
- Promoter Regions, Genetic
- Photochemical Processes
- Light
- Humans
- Hela Cells
Citation
Published In
DOI
EISSN
ISSN
Publication Date
Volume
Start / End Page
Related Subject Headings
- Zinc Fingers
- Transcriptional Activation
- Transcription, Genetic
- Transcription Factors
- Protein Engineering
- Promoter Regions, Genetic
- Photochemical Processes
- Light
- Humans
- Hela Cells