title: Strategy for naturelike designer transcription factors with reduced toxicity.
abstract: For clinical applications, the biological functions of DNA-binding proteins require that they interact with their target binding site with high affinity and specificity. Advances in randomized production and target-oriented selection of engineered artificial DNA-binding domains incited a rapidly expanding field of designer transcription factors (TFs). Engineered transcription factors are used in zinc-finger nuclease (ZFN) technology that allows targeted genome editing. Zinc-finger-binding domains fabricated by modular assembly display an unexpectedly high failure rate having either a lack of activity as ZFNs in human cells or activity at "off-targetâ binding sites on the human genome causing cell death. To address these shortcomings, we created new binding domains using a targeted modification strategy. We produced two SP1 mutants by exchanging amino acid residues in the alpha-helical region of the transcription factor SP1. We identified their best target binding sites and searched the NCBI HuRef genome for matches of the nine-base-pair consensus binding site of SP1 and the best binding sites of its mutants. Our research concludes that we can alter the binding preference of existing zinc-finger domains without altering its biological functionalities.
authors.affiliation: University of Bridgeport, Bridgeport.
authors.forename: Christian
authors.initials: C
authors.lastname: Bach
authors.affiliation: University of Bridgeport, Bridgeport.
authors.forename: Prabir K
authors.initials: PK
authors.lastname: Patra
authors.affiliation: University of Bridgeport, Bridgeport.
authors.forename: Jani M
authors.initials: JM
authors.lastname: Pallis
authors.affiliation: Bard High School, Queens.
authors.forename: William B
authors.initials: WB
authors.lastname: Sherman
authors.affiliation: University of Bridgeport, Bridgeport.
authors.forename: Hassan
authors.initials: H
authors.lastname: Bajwa
mesh_terms: D040761:Biomimetic Materials / Q000138:chemical synthesis*; D032701:Biomimetics / Q000379:methods*; D004268:DNA-Binding Proteins / Q000138:chemical synthesis* / Q000633:toxicity*; D015195:Drug Design; D014157:Transcription Factors / Q000138:chemical synthesis* / Q000235:genetics / Q000633:toxicity*; D016335:Zinc Fingers
chemical_list: D004268:DNA-Binding Proteins; D014157:Transcription Factors