Protein and RNA engineering to customize microbial molecular reporting

  • Gredell J
  • Frei C
  • Cirino P
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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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  • Joseph A. Gredell

  • Christopher S. Frei

  • Patrick C. Cirino

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