Team:NYMU-Taipei
Welcome to NYMU-Taipei team 2016!
Here is our preliminary team project descriptions!
Background of our Project
This year, we are focusing on the entomopathogenic fungus Metarhizium anisopliae, which is currently used as a biological control agent for different arthropods. However, its uses has been limited by poor efficacy. That is why many experts genetically engineered various M. anisopliae strains in order to improve its virulence. While genetic engineering is a great way to improve upon what mother nature has given us, biosafety has always been a big problem with genetic engineered organisms. For this reason, we hope to build a biosafety system, which includes one optogenetic module and one kill switch, to limit the persistence of our genetically engineered fungus in the environment.
Details of our project
In our biosafety system, our design consists of an optogenetic module and a kill switch. We used the optogenetic system VP-EL222 which containing the VP-EL222 genes and a promoter that activated by the VP-EL222 proteins blue light inducible dimerization and DNA binding. We only want the kill switch to be induced when the fungi have already killed the insect. To achieve this, we put VP-EL222 under the control of the hemolymph-induced promoter, Pmcl1. This allows the production of VP-EL222 proteins in the darkness of the insects’ interior. The fungus will breach the the dead host’s cuticle from inside after killing its host and expose to blue light, then lethality can be induced by built-in killswitch, which will be activated when the light-inducible promoter drives the expression of the proteins in the kill switch system due to VP-EL222 dimerization and the DNA binding.
When it comes to the kill switch, we employ a simple, versatile, and filamentous-fungi-specific CRISPR/Cas9 system developed by the authors of the 2015 paper “A CRISPR-Cas9 System for Genetic Engineering of Filamentous Fungi.” Because virtually any gene can be targeted by RNA-guided Streptococcus pyogenes Cas9, we target several genes that could disrupt the life cycle and reduce the survivability of the genetic engineered M. anisopliae. Two of our target genes, MrPHR1 and MrPHR2, encodes photolyases in Metarhizium. Removing them will seize the production of UV-induced cyclobutane pyrimidine dimers (CPD) and pyrimidine (6-4) photoproducts [(6-4) PPs]. The effect is supported by 2012 paper “Enhanced UV Resistance and Improved Killing of Malaria Mosquitoes by Photolyase Transgenic Entomopathogenic Fungi” which shows that deleting native photolyase genes will strictly contain M. robertsii to areas protected from sunlight, alleviating safety concerns that transgenic hypervirulent Metarhizium spp. (We will blast the same genes in the M.anisopliae strain’s genome.)
We have also chosen some other essential genes for M.anisopliae as our potential target genes. The repetible DNA regions in these essential genes will serve as the sgRNA template allowing the Cas9 to mutate many positions within the gene with high off-target activity, differing from mostly other CRISPR researches insisting Cas9 must have high on-target activity, resulting in the transformed M.anisopliae that will easily die under blue light.
What NYMU team has been working on so far?
We are still waiting for our Metarhizium anisopliae ARSEF549 which we booked from ARSEF, US. Thus, we conduct our experiences in E.coli. now. We have already completed the design of our system. However, we can't sure whether will our Metarhizium arrives on time or not. All we can do is to prepare all the other thing and as soon as the Metarhuzium arrived we can really start our project!
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.
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!
Tips
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 2016 calendar
- Have lots of fun!
Inspiration
You can also view other team wikis for inspiration! Here are some examples:
Uploading pictures and files
You can upload your pictures and files to the iGEM 2016 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 Team: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!)
