With the advent of toehold switch RNA elements and the invention of functional fibers with electrical and optical properties, more complex biosensing systems have emerged. Currently. Toehold switch-based biosensors have been developed to enable inexpensive, paper-based, cell-free visual diagnostics. In theory, the toehold switch can detect almost any RNA sequence. However, the sensitivity of the sensor decreased when longer mRNAs were detected. This may be due to the secondary structure of long mRNAs making them inaccessible, resulting in reduced sensor sensitivity. At CD BioSciences, we introduced "Helper RNAs" into a toehold switch-based biosensor to improve its sensitivity to detect long mRNAs.
Synthetic biology utilizes custom biological circuit technology to design modular biosensors, logic circuits, and output devices to achieve new control over biological systems. Similar to other toehold switch biosensor development services, in this development service, we also need to design a specific toehold switch RNA based on the target mRNA, and construct the RNA element and other reagents such as isothermal amplification enzymes into a gene circuit. The difference is that in this development service, we introduce helper RNAs that target several tens of nt upstream and downstream of the target RNA sequence. Applying helper RNAs around the target sequence helps the mRNA open up its secondary structure, making it easier to trigger the toehold switch to increase the sensitivity of the sensor.
Our toehold switch sensor with the helper RNAs development service mainly includes the following steps.
1. Synthesis of toehold switch. First, we need to determine the target sequence of the target and use machine learning algorithms to design and optimize the toehold switch, and finally synthesize the toehold switch under our high-end laboratory conditions.
2. Construction of helper RNAs. According to the target RNA sequence of about 30 nt upstream and/or downstream of the target sequence, helper RNAs that can bind to it are designed to unravel the local structure formed by the target RNA sequence and adjacent RNAs to weaken the secondary structure of the target mRNA. stability.
3. Construction of gene circuits. Mix the toehold switch RNA and helper RNAs designed in the first two steps with various enzymes and other reagents to construct a biological circuit. If a cell-free system is used, the cell-free system also needs to be constructed.
4. The constructed gene circuit is connected with output devices, etc., and finally, a biosensor based on the toehold switch with helper RNAs is constructed.
As a biotechnology company representing the international level, CD BioSciences relies on the toehold switches technology platform and an experienced team of experts to provide researchers engaged in biosensor development with customized toehold switch sensors with the helper RNAs development service. If you are interested in this service of ours, please contact us directly.