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Design and Synthesis of Artificial Zinc Finger Proteins

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Creative Biolabs offers custom design, synthesis and production services of Artificial Zinc Finger Proteins for various protein engineering studies.

Artificial zinc finger proteins can bind with DNA, and are novel tools for biochemical and molecular biological investigations and have important applications in protein engineering researches. In particular, the (Cys)2(His)2-type zinc finger motif comprises the most common class of DNA-binding proteins across all biological systems, and offers a versatile and potential framework for the design and manipulation of new DNA-binding proteins. Specially, the creation of new artificial zinc finger proteins with novel DNA-binding properties (e.g., long-DNA recognition, DNA bending, and AT-rich sequence recognition) is expected to be a key focus of zinc finger protein research. Herein, new strategies for the design of multi-zinc finger proteins for the recognition of a target DNA sequence, a DNA-bending zinc fin- ger protein, a (His)4-type zinc finger protein, and an AT-recognizing zinc finger protein are described based on recent experimental results.

Being antiparallel to the primary interacting strand of DNA, the zinc finger structure contains a repeated sequence and a specific linker. The DNA-binding residues are localized within its α-helix facing the major groove, and can recognize and make 1:1 contacts with three contiguous base-pairs of a DNA sequence. Specifically, three fingers of the (Cys)2(His)2-type zinc finger protein, Sp1, can bind to GC-rich sequences, whereas three fingers (fingers 4–6) of the six (Cys)2(His)2-type zinc finger protein, CF2-II, are binding to AT-rich sequences. α-helix substitution between those two types of zinc finger proteins gives rise to novel zinc finger protein binding to AT-rich sequences. Detailed understanding of the structure features and corresponding binding preferences can lead to greater flexibility of zinc finger engineering.

Creative Biolabs focuses on the strategies and services for the design and preparation of novel artificial zinc finger proteins with DNA-binding properties. We can offer custom protein engineering and production services including but not limited to:

  1. Nine-zinc finger proteins
  2. DNA-bending zinc finger proteins
  3. (His)4-type zinc finger proteins
  4. (Cys)2(His)2-type zinc finger proteins
  5. AT-recognizing zinc finger proteins

Figure 1. Structure of zinc finger proteins (light brown) in complex with target DNA (gray) (PDB ID: 2I13). (Charles A. Gersbach et al., 2014)

List of References:
Charles A. Gersbach, Thomas Gaj. Synthetic Zinc Finger Proteins: The Advent of Targeted Gene Regulation and Genome Modification Technologies. American Chemical Society 2014 47: 2309−2318.


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