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Related Concept Videos

Combinatorial Gene Control02:33

Combinatorial Gene Control

Combinatorial gene control is the synergistic action of several transcriptional factors to regulate the expression of a single gene. The absence of one or more of these factors may lead to a significant difference in the level of gene expression or repression.
The expression of more than 30,000 genes is controlled by approximately 2000-3000 transcription factors. This is possible because a single transcription factor can recognize more than one regulatory sequence. The specificity in gene...

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Related Experiment Video

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Direct Protein Delivery to Mammalian Cells Using Cell-permeable Cys2-His2 Zinc-finger Domains
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Published on: March 25, 2015

A tunable zinc finger-based framework for Boolean logic computation in mammalian cells.

Jason J Lohmueller1, Thomas Z Armel, Pamela A Silver

  • 1Department of Systems Biology, Harvard University, Boston, MA 02115, USA.

Nucleic Acids Research
|February 11, 2012
PubMed
Summary

Researchers created artificial zinc finger transcription factors (ZF-TFs) for molecular computation in mammalian cells. This synthetic biology platform enables precise control of cellular behavior for advanced therapies and diagnostics.

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Area of Science:

  • Synthetic biology
  • Molecular computation
  • Mammalian cell engineering

Background:

  • Cell-based therapies and diagnostics require sophisticated molecular-level computation.
  • Artificial transcription factors offer a programmable platform for gene circuit construction.

Purpose of the Study:

  • To develop a Boolean logic framework using artificial Cys(2)-His(2) zinc finger transcription factors (ZF-TFs) as computing elements.
  • To establish a method for tuning ZF-TF response and demonstrate their use in synthetic gene circuits.

Main Methods:

  • Designed and generated artificial ZF-TFs (activators and repressors).
  • Tuned ZF-TF response by fusing them to leucine zipper homodimerization domains.
  • Implemented Boolean logic gates (OR, NOR, AND, NAND) using hybrid promoters and split intein-mediated protein splicing.

Main Results:

  • Developed 15 transcriptional activators with 2- to 463-fold induction.
  • Developed 15 transcriptional repressors with 1.3- to 16-fold repression.
  • Successfully computed complex logic functions using the engineered ZF-TFs and split intein system.

Conclusions:

  • Created a robust platform for mammalian synthetic gene circuits using engineered ZF-TFs.
  • Demonstrated the potential for precise modulation of cellular behavior through molecular computation.
  • Enabled a new wave of sophisticated cell-based therapies and diagnostics.