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.
Department of Biomedical Engineering, Duke University, Room 136 Hudson Hall, 90281, Durham, NC, 27708-0281, USA.
Lauren R
LR
Polstein
Charles A
CA
Gersbach
D029681:Arabidopsis Proteins / Q000096:biosynthesis / Q000235:genetics; D001483:Base Sequence; D003001:Cloning, Molecular; D003587:Cytomegalovirus / Q000235:genetics; D017930:Genes, Reporter; D005814:Genes, Viral; D049452:Green Fluorescent Proteins / Q000096:biosynthesis / Q000235:genetics; D057809:HEK293 Cells; D006367:HeLa Cells; D006801:Humans; D008027:Light; D055668:Photochemical Processes; D011401:Promoter Regions, Genetic; D015202:Protein Engineering; D014157:Transcription Factors / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics*; D014158:Transcription, Genetic / Q000528:radiation effects; D015533:Transcriptional Activation / Q000528:radiation effects*; D016335:Zinc Fingers
D029681:Arabidopsis Proteins; C410504:FKF1 protein, Arabidopsis; C121183:GI protein, Arabidopsis; D014157:Transcription Factors; D049452:Green Fluorescent Proteins