CtrlC - Controlling Plasmids Copy Number

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Plasmid Applications

Plasmids are small circular molecules of DNA, which can most commonly be found residing in bacteria, and to a lesser extent in other kingdoms of life. In bacteria, plasmids can be found separated from the bacterial chromosome, which goes along with their inert ability to replicate independently from it. They play crucial roles in any scientific research and biotechnology, serving as essential tools in gene cloning, recombinant protein synthesis, genetic editing, and complex genetic circuits.

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Plasmid Copy Number's Impact on mRNA and Protein Levels

One critical characteristic of plasmids is their copy number, which denotes the quantity of plasmid copies within a host cell. But the quantity of the plasmid isn't the only thing affected by copy number, as copy number increases so does the concentration of the mRNA that is transcribed from the plasmid which in turn increases the level of proteins that are translated from said mRNA.

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Balancing Plasmid Numbers: A Chocolate Factory Analogy

Think of cells like a chocolate factory with its workforce (the plasmids). An excess of workers may initially seem advantageous, yet, as the old saying goes: “too much of a good thing is a bad thing”. Too many workers can impose a burden on finite resources such as finances, equipment, facilities and more potentially resulting in production issues. With that said, there is no doubt that a reduced workforce translates to reduced chocolate production.

table of different copy number rabges and their affects

Similarly, plasmids exhibit this duality. High copy number plasmids, resembling the overabundance of workers, can overburden the host cell, escalate plasmid instability, reduce cell growth rates, and compromise experimental integrity. Conversely, low copy number plasmids, like a lean workforce, limit protein production and diminish the effectiveness of genetic engineering applications. Achieving the right balance in plasmid copy number is a central aspect of synthetic biology initiatives, striking a delicate equilibrium between resource utilization and desired outcomes.

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CtrlC - Controlling Plasmid Copy Number

Until now, scientists have been limited to choosing between high and low copy number plasmids only. However, with our groundbreaking innovation, they now have access to a wide spectrum of plasmid copy numbers, all achieved by simply modifying key parts of ColE1’s origin of replication. By offering a gradient of copy numbers, our tool allows researchers to tune the expression of their genes of interest like never before.

This innovation enables scientists to avoid cellular stress caused by excessively high copy numbers or ensure sufficient yield with moderate copy numbers. Moreover, complex genetic circuits can now be finely tuned without the need of an inducer. Our technology empowers researchers to optimize their experiments and tailor outcomes to their exact specifications.

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Our Tool

Our user-friendly and easy-to-use web tool allows you to design your ColE1-like plasmid sequences with your desired copy number in order to optimize and maximize protein yield.

This tool, with its friendly user interface (UI), enables users to input plasmid sequences or select from existing ones, producing FASTA files edited to their desired copy numbers. Moreover, it enhances user experience by providing comprehensive plasmid maps with clear annotations, facilitating data interpretation, and sharing. The tool's accessibility across various devices ensures effortless use, making it an invaluable asset for geneticists and biotechnologists

web tool screenshot