@inbook{ce4acca9115241f7b3ecdfc4d2afce55,
title = "Engineered zinc finger DNA-binding domains: Synthesis, assessment of DNA-binding affinity, and direct protein delivery to mammalian cells",
abstract = "Zinc finger proteins are the most common among families of DNA-binding transcription factors. Designer transcription factors generated by the fusion of engineered zinc finger DNA-binding domains (ZF-DBDs) to effector domains have been valuable tools for the modulation of gene expression and for targeted genome editing. However, ZF-DBDs without effector domains have also been shown to effectively modulate gene expression by competing with sequence-specific DNA-binding transcription factors. Here, we describe the methodology and provide a detailed workflow for the cloning, expression, purification, and direct cell delivery of engineered ZF-DBDs. Using this protocol, ZF-DBDs can be generated with high efficiency in less than 2 weeks. We also describe a nonradioactive method for measuring DNA binding affinity of the purified ZF-DBD proteins as well as a method for direct delivery of the purified ZF-DBDs to mammalian cells.",
keywords = "Direct protein delivery, Electrophoretic mobility shift assay, Engineered zinc finger DNA-binding domain, Modular assembly, Recombinant protein purification",
author = "Hossain, {Mir A.} and Knudson, {Isaac J.} and Shaleen Thakur and Yong Shen and Stees, {Jared R.} and Barrow, {Joeva J.} and J{\"o}rg Bungert",
note = "Publisher Copyright: {\textcopyright} Springer Science+Business Media LLC 2017.",
year = "2017",
doi = "10.1007/978-1-4939-7231-9_27",
language = "English",
series = "Methods in Molecular Biology",
publisher = "Humana Press Inc.",
pages = "361--375",
booktitle = "Methods in Molecular Biology",
}