Efficient design strategy for whole-cell and cell-free biosensors based on engineered riboswitches

Norihito Muranaka, Vandana Sharma, Yoko Nomura, Yohei Yokobayashi

Research output: Contribution to journalArticlepeer-review

17 Scopus citations

Abstract

We have applied dual genetic selection to design a bacterial riboswitch with improved sensitivity by employing a high-affinity heterologous thiamine pyrophosphate (TPP) aptamer and modified selection conditions in Escherichia coli. The cells transformed with the engineered TPP riboswitches were characterized as whole-cell biosensors. The riboswitches were further studied in cell-free translation systems where they exhibited characteristics similar to those in vivo and a shorter response time for analysis. The flexibility of the riboswitch-based biosensors to accommodate different reporter genes was also demonstrated. Tuning of biosensor characteristics in vivo enables efficient development of aptamer-based whole-cell and cell-free biosensors.

Original languageEnglish (US)
Pages (from-to)108-122
Number of pages15
JournalAnalytical Letters
Volume42
Issue number1
DOIs
StatePublished - Jan 2009

Keywords

  • Cell-free biosensor
  • Dual genetic selection
  • Riboswitch
  • Whole-cell biosensor

ASJC Scopus subject areas

  • Analytical Chemistry
  • Biochemistry
  • Clinical Biochemistry
  • Biochemistry, medical
  • Electrochemistry
  • Spectroscopy

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