Protein and RNA engineering to customize microbial molecular reporting

  • Gredell J
  • Frei C
  • Cirino P
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Abstract

Nature takes advantage of the malleability of protein and RNA sequence and structure to employ these macromolecules as molecular reporters whose conformation and functional roles depend on the presence of a specific ligand (an "effector" molecule). By following nature's example, ligand-responsive proteins and RNA molecules are now routinely engineered and incorporated into customized molecular reporting systems (biosensors). Microbial small-molecule biosensors and endogenous molecular reporters based on these sensing components find a variety of applications that include high-throughput screening of biosynthesis libraries, environmental monitoring, and novel gene regulation in synthetic biology. Here, we review recent advances in engineering small-molecule recognition by proteins and RNA and in coupling in vivo ligand binding to reporter-gene expression or to allosteric activation of a protein conferring a detectable phenotype. Emphasis is placed on microbial screening systems that serve as molecular reporters and facilitate engineering the ligand-binding component to recognize new molecules.

Author-supplied keywords

  • Aptamer
  • Directed evolution
  • Molecular recognition
  • Regulatory protein
  • Riboswitch

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Authors

  • Joseph A. Gredell

  • Christopher S. Frei

  • Patrick C. Cirino

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