A Miniaturized Escherichia coli Green Light Sensor with High Dynamic Range

作者：Ong Nicholas T; Tabor Jeffrey J^*

来源：ChemBioChem, 2018, 19(12): 1255-1258.

DOI：10.1002/cbic.201800007

• gene expression
• optogenetics
• photoreceptors
• synthetic biology

Genetically engineered photoreceptors enable unrivaled control over gene expression. Previously, we ported the Synechocystis PCC 6803 CcaSR two-component system, which is activated by green light and deactivated by red, into Escherichia coli, resulting in a sensor with a sixfold dynamic range. Later, we optimized pathway protein expression levels and the output promoter sequence to decrease transcriptional leakiness and to increase the dynamic range to approximately 120-fold. These CcaSR v1.0 and v2.0 systems have been used for precise quantitative, temporal, and spatial control of gene expression for a variety of applications. Recently, other workers deleted two PAS domains of unknown function from the CcaS sensor histidine kinase in a system similar to CcaSR v1.0. Here we apply these deletions to CcaSR v2.0, resulting in a v3.0 light sensor with an output four times less leaky and a dynamic range of nearly 600-fold. We demonstrate that the PAS domain deletions have no deleterious effect on CcaSR green light sensitivity or response dynamics. CcaSR v3.0 is the best-performing engineered bacterial green light sensor available, and should have broad applications in fundamental and synthetic biology studies.

• 出版日期2018-6-18