Team:MIT/Attributions

• BACKGROUND
• EXPERIMENTS
  Synthetic Promoters microRNA Recombinase Proof of Concept Collaborations
• FUTURE WORK
  Circuit Design Clinical Applications
• HUMAN PRACTICES
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• NOTEBOOK
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• TEAM

Attributions

We'd like to thank everyone who helped us this year to make iGEM not only possible but extremely fun! This has been a great experience and we couldn't have done it without help!

Promoter Engineering Subgroup Attributions

All estrogen and progesterone sensing constructs were designed and created by the members of our subgroup. Entry vectors for constitutive promoters (hEF1a), repressors (TAL14, BM3R1,TAL21), fluorescent genes (eYFP, BFP, etc.), and activators were provided by the Weiss Lab. All experiments on our promoters were performed by members of our subgroup and conceived by us the students. The student on our team who proposed the minCMV architecture with upstream binding sites for the ER and PR was inspired by a class he had taken with Prof. Ron Weiss, Biological Circuit Engineering Laboratory, where he had done some basic promoter engineering work in yeast. The cells we sought to test our promoters with (MCF7, ISH, tHESC) were not available in our host lab (the Weiss Lab), so we initiated a collaboration with the Grifith Lab at MIT, affiliated with the Center for Gynepathology Research. Christi Cook and Linda Stockdale of the Griffith Lab provided us with the cells, as well as best practices for culturing and stimulating them. Prof. Linda Griffith and Prof. Ron Weiss advised us during lab meetings where we would present our data. Postdoc Brian Teague of the Weiss Lab was our direct supervisor in case we ran into any difficulties with our labwork; he also helped us initially optimize the transfection of tHESC using electroporation when we ran into difficulties with cationic lipid transfection.

miRNA Sensing Subgroup Attributions

All experiments were performed by members of our subgroup and conceived by us the students. All cloning was performed by members of our subgroup. The design, creation, and testing of pDest_mCherry was spearheaded by us after initial inspiration from Weiss Lab postdoc Brian Teague. Entry vectors for constitutive promoters (hEF1a), repressors (TAL14, BM3R1,TAL21), fluorescent genes (eYFP, BFP, etc.), and activators were provided by the Weiss Lab. miRNA sensors were provided by graduate student Jeremy Gam. tHESC cells were provided by a new collaboration we initiated with Prof. Griffith in the Weiss Lab. Postdoc Brian Teague of the Weiss Lab was our direct supervisor in case we ran into any difficulties with our labwork; he also helped us initially optimize the transfection of tHESC using electroporation when we ran into difficulties with cationic lipid transfection.

Recombinase Subgroup Attributions

All experiments were performed by members of our subgroup and conceived by us the students. All cloning was performed by members of our subgroup. Entry vectors for constitutive promoters (hEF1a), repressors (TAL14, BM3R1,TAL21), fluorescent genes (eYFP, BFP, etc.), and activators (VgEcr, rtTA) were provided by the Weiss Lab. Sequences for the recombinases with nuclear localization sequences were obtained through a collaboration with the Boston University Wet Lab iGEM team. Postdoc Brian Teague of the Weiss Lab was our direct supervisor in case we ran into any difficulties with our labwork.

The Weiss Lab - The Center for Synthetic Biology

Brian Teague

Mentored us throughout the process of developing our own project and coached us in the lab skills we needed to implement it.

Jeremy Gam

Provided us with the miRNA sensors and advice on learning about and testing for different miRNA expression levels.

Ron Weiss

Provided us with resources from the lab and feedback on the direction of the project.

The Griffith Lab - Center for Gynepathology Research

Linda Stockdale

Provided us with splits of tHESC, ISH, and MCF7 as well as estrogen and progesterone.

Christi Cook

Advising us on dilution protocols and best practice cell culture techniques.

Linda Griffith

Provided us with resources from the lab and feedback on the direction of the project.