Mini Photobioreactors for in Vivo Real-Time Characterization and Evolutionary Tuning of Bacterial Optogenetic Circuit

ACS Synthetic Biology
Hsinkai Wang, Ya-Tang Yang

Abstract

The current standard protocols for characterizing the optogenetic circuit of bacterial cells using flow cytometry in light tubes and light exposure of culture plates are tedious, labor-intensive, and cumbersome. In this work, we engineer a bioreactor with working volume of ∼10 mL for in vivo real-time optogenetic characterization of E. coli with a CcaS-CcaR light-sensing system. In the bioreactor, optical density measurements, reporter protein fluorescence detection, and light input stimuli are provided by four light-emitting diode sources and two photodetectors. Once calibrated, the device can cultivate microbial cells and record their growth and gene expression without human intervention. We measure gene expression during cell growth with different organic substrates (glucose, succinate, acetate, pyruvate) as carbon sources in minimal medium and demonstrate evolutionary tuning of the optogenetic circuit by serial dilution passages.

References

Sep 10, 2002·Nature Biotechnology·Sae Shimizu-SatoPeter H Quail
Nov 25, 2005·Nature·Anselm LevskayaChristopher A Voigt
Sep 1, 2006·Applied Microbiology and Biotechnology·Oleg Paliy, Thusitha S Gunasekera
Sep 4, 2007·Journal of Molecular Biology·Andreas Möglich, Keith Moffat
Dec 27, 2008·Journal of Molecular Biology·Andreas MöglichKeith Moffat
Mar 10, 2010·Nature Biotechnology·Jeffrey D OrthBernhard Ø Palsson
Nov 3, 2010·Nature Methods·Matthew J KennedyChandra L Tucker
Apr 2, 2011·Science·Luca RizziniRoman Ulm
Jan 17, 2012·Journal of Molecular Biology·Robert OhlendorfAndreas Möglich
May 11, 2012·The Journal of Biological Chemistry·John M ChristieElizabeth D Getzoff
Aug 10, 2013·Journal of Molecular Biology·Eric A DavidsonTravis S Bayer
Jun 14, 2014·ACS Synthetic Biology·Min-Hyung Ryu, Mark Gomelsky
Jul 19, 2014·ACS Synthetic Biology·Chris N TakahashiEric Klavins
Sep 25, 2014·ACS Synthetic Biology·Sebastian R SchmidlJeffrey J Tabor
Nov 7, 2014·Photochemical & Photobiological Sciences : Official Journal of the European Photochemistry Association and the European Society for Photobiology·Florian RichterAndreas Möglich
Nov 3, 2016·Scientific Reports·Karl P GerhardtJeffrey J Tabor

❮ Previous
Next ❯

Citations

Nov 10, 2018·Nature Reviews. Microbiology·Lisa B Wiltbank, David M Kehoe
Apr 25, 2020·Biotechnology Progress·Ding-Shun LinYa-Tang Yang
Nov 19, 2021·FEMS Microbiology Reviews·Florian Lindner, Andreas Diepold

❮ Previous
Next ❯

Related Concepts

Trending Feeds

COVID-19

Coronaviruses encompass a large family of viruses that cause the common cold as well as more serious diseases, such as the ongoing outbreak of coronavirus disease 2019 (COVID-19; formally known as 2019-nCoV). Coronaviruses can spread from animals to humans; symptoms include fever, cough, shortness of breath, and breathing difficulties; in more severe cases, infection can lead to death. This feed covers recent research on COVID-19.

Blastomycosis

Blastomycosis fungal infections spread through inhaling Blastomyces dermatitidis spores. Discover the latest research on blastomycosis fungal infections here.

Nuclear Pore Complex in ALS/FTD

Alterations in nucleocytoplasmic transport, controlled by the nuclear pore complex, may be involved in the pathomechanism underlying multiple neurodegenerative diseases including Amyotrophic Lateral Sclerosis and Frontotemporal Dementia. Here is the latest research on the nuclear pore complex in ALS and FTD.

Applications of Molecular Barcoding

The concept of molecular barcoding is that each original DNA or RNA molecule is attached to a unique sequence barcode. Sequence reads having different barcodes represent different original molecules, while sequence reads having the same barcode are results of PCR duplication from one original molecule. Discover the latest research on molecular barcoding here.

Chronic Fatigue Syndrome

Chronic fatigue syndrome is a disease characterized by unexplained disabling fatigue; the pathology of which is incompletely understood. Discover the latest research on chronic fatigue syndrome here.

Evolution of Pluripotency

Pluripotency refers to the ability of a cell to develop into three primary germ cell layers of the embryo. This feed focuses on the mechanisms that underlie the evolution of pluripotency. Here is the latest research.

Position Effect Variegation

Position Effect Variagation occurs when a gene is inactivated due to its positioning near heterochromatic regions within a chromosome. Discover the latest research on Position Effect Variagation here.

STING Receptor Agonists

Stimulator of IFN genes (STING) are a group of transmembrane proteins that are involved in the induction of type I interferon that is important in the innate immune response. The stimulation of STING has been an active area of research in the treatment of cancer and infectious diseases. Here is the latest research on STING receptor agonists.

Microbicide

Microbicides are products that can be applied to vaginal or rectal mucosal surfaces with the goal of preventing, or at least significantly reducing, the transmission of sexually transmitted infections. Here is the latest research on microbicides.