Light & Tet Operon Dually Controlled System

A Light & Tet Operon Dually Controlled System (Light/Tet System for short) construction service is now available at Creative BioMart. The system integrates the advantages of light and chemical induction, and can accurately regulate the expression of target genes at single-cell resolution.

Tet-OFF/ON System and CRY2/CIB1 Switch

Tet-OFF/ON system is the most commonly used chemical regulatory system in mammalian cells. In this system, small molecules like tetracycline (Tet) or doxycycline (Dox) are used to treat the expression of foreign genes in the target cell. The Tet-OFF system activates downstream gene expression in the absence of Dox, while the Tet-ON system activates it in the presence of Dox.

Cryptochromes are flavoprotein photoreceptors that first identified in Arabidopsis thaliana, where they regulate growth and developmental processes. It has been found in bacteria, higher plants, and animals. The blue light-responsive cryptochrome 2/cryptochrome-interacting basic-helix-loop-helix 1 (CRY2/CIB1) system has become a popular optogenetic tool for gene expression control.

What We Can Do For You?

Light/Tet System is consist of three parts which are exist independently

Plasmid 1——The Tet-OFF/ON plasmid that harbors theTetracycline reaction element (TRE) and the downstream GOI.

Plasmid 2——Tet repressor (TetR) that fused with CIB1.

Plasmid 3——Transcription activation domain of RELA that fused with CRY2.

Light/Tet System for in vitro application

The system has the ability to induce light triggered gene expression in primary cultured cells and tissues. Our services include

Step	Service Description
Expression strategy design	Client provides the target sequence and expression strategy designed referring to customers' requirement.
Plasmid design	Codon optimization Clone the GOI sequence downstream the TRE
Plasmid transformation	ransfer the recombinant plasmid 1 into the host cell together with plasmid 2 and 3 by means of electroporation or AAV plasmid deliver.

In addition to the in vitro application, the system also has the ability to induce light triggered gene expression for in vivo application. Please feel free to contact us if you are interested in.

Features of the Light/Tet System

Light/Tet system has low background and high light activation multiple, and can be used in a variety of mammalian cells
By changing the applied light intensity and Dox concentration, more fine and adjustable gene expression can be realized in the system
By limiting the application of Dox in the system, light controlled gene expression can be conditionally induced in a narrow time window during long-term experiments in vitro and in vivo
The dual control function of the system is conducive to the experiment of system biology, in which the time, quantity and mode of gene expression must be strictly controlled.

Creative BioMart is always devote us to provide high-quality and satisfactory service to our customers, if you are interested in our services or have some question, please feel free to contact us or make online inquiry.

References

Yamada M, Suzuki Y, Nagasaki SC, Okuno H, Imayoshi I. Light Control of the Tet Gene Expression System in Mammalian Cells. Cell Rep. 2018 Oct 9;25(2):487-500.e6.
Yamada M, Nagasaki SC, Ozawa T, Imayoshi I. Light-mediated control of Gene expression in mammalian cells. Neurosci Res. 2020 Mar;152:66-77.