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Related Concept Videos

Protein Complexes with Interchangeable Parts01:57

Protein Complexes with Interchangeable Parts

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Groups of proteins may form a complex where each protein in this complex has a different role in the overall execution of the complex’s function. Often some of the proteins in the complex can be replaced by a closely related variant to give a complex that contains many of the same components yet is functionally distinct.
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Cooperative allosteric transitions can occur in multimeric proteins, where each subunit of the protein has its own ligand-binding site. When a ligand binds to any of these subunits, it triggers a conformational change that affects the binding sites in the other subunits; this can change the affinity of the other sites for their respective ligands. The ability of the protein to change the shape of its binding site is attributed to the presence of a mix of flexible and stable segments in the...
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Transcriptional regulators bind to specific cis-regulatory sequences in the DNA to regulate gene transcription. These cis-regulatory sequences are very short, usually less than ten nucleotide pairs in length. The short length means that there is a high probability of the exact same sequence randomly occurring throughout the genome.  Since regulators can also bind to groups of similar sequences, this further increases the chances of random binding. Transcriptional regulators form...
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Related Experiment Video

Updated: Aug 7, 2025

Creating Highly Specific Chemically Induced Protein Dimerization Systems by Stepwise Phage Selection of a Combinatorial Single-Domain Antibody Library
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Combinatorial protein dimerization enables precise multi-input synthetic computations.

Adrian Bertschi1, Pengli Wang1, Silvia Galvan1

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Researchers engineered bacterial transcription factors (TFs) to create novel gene switches in mammalian cells. This modular approach enables complex logic gates and new inducible systems for synthetic biology applications.

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Area of Science:

  • Synthetic Biology
  • Molecular Biology
  • Genetics

Background:

  • Bacterial transcription factors (TFs) with helix-turn-helix (HTH) DNA-binding domains are utilized for orthogonal transcriptional regulation in mammalian cells.
  • The modular nature of these TFs offers potential for constructing sophisticated genetic circuits.
  • Existing systems often rely on DNA-binding-dependent activation, limiting versatility.

Purpose of the Study:

  • To develop a versatile framework for multi-input logic gates using bacterial TFs.
  • To engineer novel ON-type gene switches and systems responsive to new inducers.
  • To create complex genetic logic circuits, including bandpass filters and multi-input AND/OR gates.

Main Methods:

  • Utilizing the modular structure of bacterial TFs, specifically fusing HTH domains to create dimerization-dependent activation.
  • Converting OFF-type gene switches to ON-type systems by engineering dimerization-dependent activation.
  • Combining OFF and ON modes to construct a bandpass filter.
  • Demonstrating cytosolic and extracellular dimerization.
  • Cascading pairwise fusion proteins to build multi-input AND logic gates and various 4-input 1-output AND/OR configurations.

Main Results:

  • Established that HTH domains alone are sufficient for DNA binding in some TFs.
  • Successfully converted DNA-binding-dependent activation to dimerization-dependent activation, enabling ON-type gene switches.
  • Created new mammalian gene switches responsive to novel inducers.
  • Developed a compact, high-performance bandpass filter by integrating OFF and ON modes.
  • Demonstrated robust multi-input AND logic gates by cascading up to five pairwise fusion proteins.
  • Configured diverse 4-input 1-output AND and OR logic gates using combinations of pairwise fusion proteins.

Conclusions:

  • A novel framework for multi-input logic gates was established using bacterial TFs and inducible protein-protein interactions.
  • Engineered dimerization-dependent activation provides a versatile strategy for creating ON-type gene switches and expanding inducer responsiveness.
  • The developed system allows for the construction of complex genetic circuits, including bandpass filters and combinatorial logic gates, with high performance and robustness.