For better understanding of our software package please visit first our Model page:
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Software
Overview
During the process of modeling our project, the team’s dry lab coded a software tool which designs and analyzes the sequences of toehold switches based on the inputs given to it (trigger, RBS, GFP, number of paired bases). The software tool utilizes Nupack, ViennaRNA (documentation in our Model's Page) and Python (version at least 3.6) to build and to characterize the behavior of the molecules. As handling the modeling in different types of computer software (Linux, Ubuntu, Windows) is quite challenging and a great spectrum of the values calculated depends on the software used for the analysis part, we decided to build the tool so that it is applicable in any case. Therefore, Nupack should be used in Linux or Ubuntu software and ViennaRNA in Windows.
For better understanding of our software package please visit first our Model page:
For better understanding of our software package please visit first our Model page:
Design and Analysis
| Nupack Design
Nupack design utilizes Nupack’s tools to build the sequences. The desired structure as defined in "Model" is created via Nupack’s functions.
| Standard Design
This particular process of design of the toehold sequence is based on the bibliography (as analyzed in "Model"). For standard design, the following apply:
- Part_1 = "GGG" + unpaired bases of complementary_part
- Part_2 = paired bases of complementary_part + "GUA" + "GUGUGU"
- Part_3 = RBS
- Part_4 = "ACACAC" + "AUG" + bases complementary to paired bases of complementary_part
- Part_5 = Linker
- Part_6 = GFP
- Toehold Sequence = Part_1 + Part_2 + Part_3 + Part_4 + Part_5 + Part_6
| Nupack Analysis
For the designed sequences the following values are calculated via Nupack’s analysis functions:- ~ΔG free energy (toehold switch)
- ~ΔG binding (miRNA – Toehold)
- ~ΔG from RBS to Linker
- ~Pair probability matrix
- ~Sampled structures
- ~Mfe structures
- ~Partition function
| ViennaRNA Analysis
For the designed sequences the following values are calculated via Nupack’s analysis functions:- ~ΔG free energy (toehold switch)
- ~ΔG binding (miRNA – Toehold)
- ~ΔG from RBS to Linker
- ~Perfect matches
- ~Concentration dependency plot
- ~Frequency of the mfe structure
- ~PostScript images(.ps files) that depicts the complex of the toehold switch and miRNA binding
- ~Scalable Vector Graphics(.svg files) that depicts the secondary structure of every toehold switch sequence
Software Table
For more details on how to use the software tool please read the manuals: