Regulation of Gene Expression Using Toehold Switches

Regulation of Gene Expression Using Toehold Switches

Regulation of Gene Expression Using Toehold Switches

Gene expression regulation refers to the regulation and control of the content and activity of proteins in the body through the regulation system in the organism, so that it is in an orderly state at a specific time and space, and appears with a certain intensity so that the body can adapt to the environment, maintains its growth and proliferation, as well as the development and differentiation of individuals. Gene regulation is one of the central topics of modern molecular biology research. In order to understand the laws of animal and plant development and biological functions, we will use some biotechnologies to regulate gene expression. 

Conventional Methods of Gene Expression Regulation

Gene expression can be regulated mainly from the following aspects: regulation at the transcriptional level; regulation at the mRNA processing and maturation level; regulation at the translational level. With the development of biology, gene editing technology appeared. We often use gene knock-in/gene knock-out to regulate gene expression at the DNA level, a typical way is CRISPR-Cas9. Gene knockdown is used to regulate gene expression at the mRNA level, and representative methods are siRNA, shRNA, CRISPR-CasRx, etc.

Toehold Switches in Gene Expression Regulation

Toehold switches are RNA engineering elements capable of de novo synthesis, consisting of triggering two RNA strands and a hairpin structure. One of the RNA strands contains the coding sequence of the regulated gene, the hairpin structure contains the ribosome binding site, and at least the codon AUG is located upstream of this sequence. Toehold switches can respond to arbitrary RNAs, making them RNA-to-protein signal transponders. Based on this ability, scientists integrated toehold switches into the genome, allowing them to sense cellular endogenous RNAs and regulate the expression of endogenous genes. Some scholars have used λ Red recombination to integrate toehold switches upstream of the target gene, thereby inhibiting the translation of the target gene. But when the trigger RNA appeared, the target gene continued to be expressed. At CD BioSciences, leveraging the established toehold switches technology platform, we provide you with design services for RNA engineering elements of toehold switches capable of regulating the expression of specific genes.

The Services We Can Provide Based on Toehold Switches

Toehold switches are codable engineered elements that can be integrated into the genome to regulate the translation of endogenous genes. As a biotechnology company with a professional toehold switches technology platform, CD BioSciences can provide the following services for specific genes for teams engaged in biosynthesis research and molecular biology research.

Application of Toehold Switches to Regulate Gene Expression

  • Study of gene function
  • Study of protein function
  • Study of protein interactions
  • Specific protein synthesis platforms

Advantages of Toehold Switches in Gene Expression Regulation

  • No modification of trigger RNA is required.
  • Ability to control the expression of any gene.
  • The reaction thermodynamics can be improved by modifying the RNA structure of toehold switches.
  • The dynamic range of this element can be increased by an unpaired region upstream of the RBA binding site.

As a biotechnology company focusing on cutting-edge technologies, CD BioSciences can provide design, optimization, and synthesis services for toehold switches in applications related to gene expression regulation. We are confident that we can make you satisfied, if you are interested, please feel free to contact us.


  1. Green, A.A.; et al. Toehold switches: de-novo-designed regulators of gene expression. Cell. 2014, 159(4): 925-939.
  2. Kim, S.J.; et al. Modulating Responses of Toehold Switches by an Inhibitory Hairpin. ACS Synth Biol. 2019, 8(3): 601-605.
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