Light-inducible gene regulation with engineered zinc finger proteins

Methods Mol Biol. 2014:1148:89-107. doi: 10.1007/978-1-4939-0470-9_7.

Abstract

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.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Arabidopsis Proteins / biosynthesis
  • Arabidopsis Proteins / genetics
  • Base Sequence
  • Cloning, Molecular
  • Cytomegalovirus / genetics
  • Genes, Reporter
  • Genes, Viral
  • Green Fluorescent Proteins / biosynthesis
  • Green Fluorescent Proteins / genetics
  • HEK293 Cells
  • HeLa Cells
  • Humans
  • Light
  • Photochemical Processes
  • Promoter Regions, Genetic
  • Protein Engineering
  • Transcription Factors / biosynthesis
  • Transcription Factors / chemistry
  • Transcription Factors / genetics*
  • Transcription, Genetic / radiation effects
  • Transcriptional Activation / radiation effects*
  • Zinc Fingers

Substances

  • Arabidopsis Proteins
  • FKF1 protein, Arabidopsis
  • GI protein, Arabidopsis
  • Transcription Factors
  • Green Fluorescent Proteins