Ratiometric Gas Reporting: A Nondisruptive Approach to Monitor Gene Expression in Soils

Hsiao Ying Cheng; Caroline A. Masiello; Ilenne Del Valle; Xiaodong Gao; George N. Bennett; Jonathan J. Silberg

doi:10.1021/acssynbio.7b00405

Ratiometric Gas Reporting: A Nondisruptive Approach to Monitor Gene Expression in Soils

Hsiao Ying Cheng, Caroline A. Masiello, Ilenne Del Valle, Xiaodong Gao, George N. Bennett, Jonathan J. Silberg

Research output: Contribution to journal › Article › peer-review

20 Scopus citations

Abstract

Fluorescent proteins are ubiquitous tools that are used to monitor the dynamic functions of natural and synthetic genetic circuits. However, these visual reporters can only be used in transparent settings, a limitation that complicates nondisruptive measurements of gene expression within many matrices, such as soils and sediments. We describe a new ratiometric gas reporting method for nondisruptively monitoring gene expression within hard-to-image environmental matrices. With this approach, C₂H₄ is continuously synthesized by ethylene forming enzyme to provide information on viable cell number, and CH₃Br is conditionally synthesized by placing a methyl halide transferase gene under the control of a conditional promoter. We show that ratiometric gas reporting enables the creation of Escherichia coli biosensors that report on acylhomoserine lactone (AHL) autoinducers used for quorum sensing by Gram-negative bacteria. Using these biosensors, we find that an agricultural soil decreases the bioavailable concentration of a long-chain AHL up to 100-fold. We also demonstrate that these biosensors can be used in soil to nondisruptively monitor AHLs synthesized by Rhizobium leguminosarum and degraded by Bacillus thuringiensis. Finally, we show that this new reporting approach can be used in Shewanella oneidensis, a bacterium that lives in sediments.

Original language	English
Pages (from-to)	903-911
Number of pages	9
Journal	ACS Synthetic Biology
Volume	7
Issue number	3
DOIs	https://doi.org/10.1021/acssynbio.7b00405
State	Published - Mar 16 2018
Externally published	Yes

Funding

We are grateful for financial support from the W. M. Keck Foundation, Rice University, and the Taiwan Ministry of Education Scholarship. Support for this research was also provided by the NSF Long-Term Ecological Research Program (DEB 1027253) at the Kellogg Biological Station and by Michigan State University AgBioResearch, and by the US DOE Office of Science (DE-FCO2-07ER64494) and Office of Energy Efficiency and Renewable Energy (DE-ACO5-76RL01830) to the DOE Great Lakes Bioenergy Research Center.

Funders	Funder number
Kellogg Biological Station
NSF Long-Term Ecological Research Program	DEB 1027253
National Science Foundation	1637653, 1027253
U.S. Department of Energy
W. M. Keck Foundation
Office of Science	DE-FCO2-07ER64494
Office of Energy Efficiency and Renewable Energy	DE-ACO5-76RL01830
Rice University
AgBioResearch, Michigan State University

Keywords

biosensor
gene expression
reporter
signaling
soil
synthetic biology

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

Access to Document

10.1021/acssynbio.7b00405

Cite this

@article{bf3784f446dd417193f46ea0f3c7c383,

title = "Ratiometric Gas Reporting: A Nondisruptive Approach to Monitor Gene Expression in Soils",

abstract = "Fluorescent proteins are ubiquitous tools that are used to monitor the dynamic functions of natural and synthetic genetic circuits. However, these visual reporters can only be used in transparent settings, a limitation that complicates nondisruptive measurements of gene expression within many matrices, such as soils and sediments. We describe a new ratiometric gas reporting method for nondisruptively monitoring gene expression within hard-to-image environmental matrices. With this approach, C2H4 is continuously synthesized by ethylene forming enzyme to provide information on viable cell number, and CH3Br is conditionally synthesized by placing a methyl halide transferase gene under the control of a conditional promoter. We show that ratiometric gas reporting enables the creation of Escherichia coli biosensors that report on acylhomoserine lactone (AHL) autoinducers used for quorum sensing by Gram-negative bacteria. Using these biosensors, we find that an agricultural soil decreases the bioavailable concentration of a long-chain AHL up to 100-fold. We also demonstrate that these biosensors can be used in soil to nondisruptively monitor AHLs synthesized by Rhizobium leguminosarum and degraded by Bacillus thuringiensis. Finally, we show that this new reporting approach can be used in Shewanella oneidensis, a bacterium that lives in sediments.",

keywords = "biosensor, gene expression, reporter, signaling, soil, synthetic biology",

author = "Cheng, {Hsiao Ying} and Masiello, {Caroline A.} and {Del Valle}, Ilenne and Xiaodong Gao and Bennett, {George N.} and Silberg, {Jonathan J.}",

note = "Publisher Copyright: {\textcopyright} 2018 American Chemical Society.",

year = "2018",

month = mar,

day = "16",

doi = "10.1021/acssynbio.7b00405",

language = "English",

volume = "7",

pages = "903--911",

journal = "ACS Synthetic Biology",

issn = "2161-5063",

publisher = "American Chemical Society",

number = "3",

}

TY - JOUR

T1 - Ratiometric Gas Reporting

T2 - A Nondisruptive Approach to Monitor Gene Expression in Soils

AU - Cheng, Hsiao Ying

AU - Masiello, Caroline A.

AU - Del Valle, Ilenne

AU - Gao, Xiaodong

AU - Bennett, George N.

AU - Silberg, Jonathan J.

PY - 2018/3/16

Y1 - 2018/3/16

N2 - Fluorescent proteins are ubiquitous tools that are used to monitor the dynamic functions of natural and synthetic genetic circuits. However, these visual reporters can only be used in transparent settings, a limitation that complicates nondisruptive measurements of gene expression within many matrices, such as soils and sediments. We describe a new ratiometric gas reporting method for nondisruptively monitoring gene expression within hard-to-image environmental matrices. With this approach, C2H4 is continuously synthesized by ethylene forming enzyme to provide information on viable cell number, and CH3Br is conditionally synthesized by placing a methyl halide transferase gene under the control of a conditional promoter. We show that ratiometric gas reporting enables the creation of Escherichia coli biosensors that report on acylhomoserine lactone (AHL) autoinducers used for quorum sensing by Gram-negative bacteria. Using these biosensors, we find that an agricultural soil decreases the bioavailable concentration of a long-chain AHL up to 100-fold. We also demonstrate that these biosensors can be used in soil to nondisruptively monitor AHLs synthesized by Rhizobium leguminosarum and degraded by Bacillus thuringiensis. Finally, we show that this new reporting approach can be used in Shewanella oneidensis, a bacterium that lives in sediments.

AB - Fluorescent proteins are ubiquitous tools that are used to monitor the dynamic functions of natural and synthetic genetic circuits. However, these visual reporters can only be used in transparent settings, a limitation that complicates nondisruptive measurements of gene expression within many matrices, such as soils and sediments. We describe a new ratiometric gas reporting method for nondisruptively monitoring gene expression within hard-to-image environmental matrices. With this approach, C2H4 is continuously synthesized by ethylene forming enzyme to provide information on viable cell number, and CH3Br is conditionally synthesized by placing a methyl halide transferase gene under the control of a conditional promoter. We show that ratiometric gas reporting enables the creation of Escherichia coli biosensors that report on acylhomoserine lactone (AHL) autoinducers used for quorum sensing by Gram-negative bacteria. Using these biosensors, we find that an agricultural soil decreases the bioavailable concentration of a long-chain AHL up to 100-fold. We also demonstrate that these biosensors can be used in soil to nondisruptively monitor AHLs synthesized by Rhizobium leguminosarum and degraded by Bacillus thuringiensis. Finally, we show that this new reporting approach can be used in Shewanella oneidensis, a bacterium that lives in sediments.

KW - biosensor

KW - gene expression

KW - reporter

KW - signaling

KW - soil

KW - synthetic biology

UR - http://www.scopus.com/inward/record.url?scp=85044040254&partnerID=8YFLogxK

U2 - 10.1021/acssynbio.7b00405

DO - 10.1021/acssynbio.7b00405

M3 - Article

C2 - 29366321

AN - SCOPUS:85044040254

SN - 2161-5063

VL - 7

SP - 903

EP - 911

JO - ACS Synthetic Biology

JF - ACS Synthetic Biology

IS - 3

ER -

Ratiometric Gas Reporting: A Nondisruptive Approach to Monitor Gene Expression in Soils

Abstract

Funding

Keywords

UN SDGs

Access to Document

Other files and links

Fingerprint

Cite this