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

Norihito Muranaka; Vandana Sharma; Yoko Nomura; Yohei Yokobayashi

doi:10.1080/00032710802568556

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

Norihito Muranaka, Vandana Sharma, Yoko Nomura, Yohei Yokobayashi

Biomedical Engineering

Research output: Contribution to journal › Article

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 language	English (US)
Pages (from-to)	108-122
Number of pages	15
Journal	Analytical Letters
Volume	42
Issue number	1
DOIs	https://doi.org/10.1080/00032710802568556
State	Published - 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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Muranaka, N., Sharma, V., Nomura, Y., & Yokobayashi, Y. (2009). Efficient design strategy for whole-cell and cell-free biosensors based on engineered riboswitches. Analytical Letters, 42(1), 108-122. https://doi.org/10.1080/00032710802568556

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