Antimicrobial resistance is a major emerging threat as reported by the WHO. Worldwide implementation of bacteriophage therapy, a 100-year old treatment employing the natural enemies of bacteria, is impeded by the lack of common manufacturing procedures which meet international quality and safety standards. Based on synthetic biology we created Phactory, a cell-free molecular assembly line for bacteriophages. We demonstrate expression of several phages including T7, MS2 and 3S at clinically relevant concentrations. Exploiting the open nature of cell-free systems, Phactory enables modular composition of bacteriophages with engineered proteins while remaining GMO-free. We developed a quality control structure utilizing state-of-the-art bioinformatics, as well as purification and encapsulation protocols. To expand our production variety while reducing cost, we optimized and engineered home-made E. Coli cell-extract. Compared to traditional manufacturing procedures, Phactory requires 2.5% of the production volume and demands no special biosafety regulations to yield bacteriophages ready for therapy.




Cell-free extract engineering

One Problem of cell extract, is the stability of linear DNA. In escherichia coli, endogenous and exogenous linear DNA are differentiated by specific proteins, such as the recombinase complex RecBCD. It recognizes genomic double strand breaks via a specific sequence of 8 bases called Chi-site and prepares the DNA for homologous repair. However, foreign DNA lacks the Chi-site and is therefore degraded by RecBCD. For experiments in cell extract expensive and inefficient inhibitors are used. Since a knockout of RecBCD is lethal, we chose to modify the enzyme to enable selective depletion after cell lysis. Therefore, we came up with several depletion strategies:

Cell free protocol optimization

The financial advantage of producing home-made cell extract for in vitro protein expression is virtually reversed by the fact that it is tedious and time-consuming, with huge batch to batch variations of the resulting product. Therefore, our objective is to optimize the protocol for self-made cell extract from E. coli, by simplifying time consuming steps while eliminating the central sources of variation during the preparation process. The focus of our effort is to determine the influence of cultivation conditions on extract quality and optimizing the cell lysis for speed, protein yield and nal product quality. By implementing novel ways of extract characterization, we aim to gain a deeper understanding of cell-free systems.

Phage engineering and phage assembly in TX/TL

Generation of a universal platform for phage engineering: Our TX-TL system is superior to conventional bacterial based phage production in the following aspects:
 - Assembly of phages in vitro
 - Exchange of phage fibers in vitro
 - Establishment of homogeneous phage batches
 - Reduction of endotoxins within phage batches We will exemplarily engineer a non lytic fluorescent T7 phage, which targets a non primary E. coli strain.

Before you start

Please read the following pages:

Styling your wiki

You may style this page as you like or you can simply leave the style as it is. You can easily keep the styling and edit the content of these default wiki pages with your project information and completely fulfill the requirement to document your project.

While you may not win Best Wiki with this styling, your team is still eligible for all other awards. This default wiki meets the requirements, it improves navigability and ease of use for visitors, and you should not feel it is necessary to style beyond what has been provided.

Uploading pictures and files

You must upload any pictures and files to the iGEM 2018 server. Remember to keep all your pictures and files within your team's namespace or at least include your team's name in the file name.

When you upload, set the "Destination Filename" to T--YourOfficialTeamName--NameOfFile.jpg. (If you don't do this, someone else might upload a different file with the same "Destination Filename", and your file would be erased!)

Wiki template information

We have created these wiki template pages to help you get started and to help you think about how your team will be evaluated. You can find a list of all the pages tied to awards here at the Pages for awards link. You must edit these pages to be evaluated for medals and awards, but ultimately the design, layout, style and all other elements of your team wiki is up to you!

Editing your wiki

On this page you can document your project, introduce your team members, document your progress and share your iGEM experience with the rest of the world!

Use WikiTools - Edit in the black menu bar to edit this page


This wiki will be your team’s first interaction with the rest of the world, so here are a few tips to help you get started:

  • State your accomplishments! Tell people what you have achieved from the start.
  • Be clear about what you are doing and how you plan to do this.
  • You have a global audience! Consider the different backgrounds that your users come from.
  • Make sure information is easy to find; nothing should be more than 3 clicks away.
  • Avoid using very small fonts and low contrast colors; information should be easy to read.
  • Start documenting your project as early as possible; don’t leave anything to the last minute before the Wiki Freeze. For a complete list of deadlines visit the iGEM 2018 calendar
  • Have lots of fun!


You can also view other team wikis for inspiration! Here are some examples: