|Title||Light-inducible gene regulation with engineered zinc finger proteins.|
|Publication Type||Journal Article|
|Year of Publication||2014|
|Authors||LR Polstein, and CA Gersbach|
|Journal||Methods in Molecular Biology (Clifton, N.J.)|
|Pagination||89 - 107|
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.
|Short Title||Methods in Molecular Biology (Clifton, N.J.)|