Difference between revisions of "Team:MIT/Experiments/Promoters"
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<img src="https://static.igem.org/mediawiki/2016/thumb/4/4f/T--MIT--KHB1177menstrual-cycle.jpeg/240px-T--MIT--KHB1177menstrual-cycle.jpeg" alt = 'menstrual cycle diagram' style='width: 250px: height = 250px; float:right;" margin: 0 1.5%;> | <img src="https://static.igem.org/mediawiki/2016/thumb/4/4f/T--MIT--KHB1177menstrual-cycle.jpeg/240px-T--MIT--KHB1177menstrual-cycle.jpeg" alt = 'menstrual cycle diagram' style='width: 250px: height = 250px; float:right;" margin: 0 1.5%;> | ||
<h2 style="text-decoration:underline; font-family: Trebuchet MS;"> Menstrual Cycle</h2> | <h2 style="text-decoration:underline; font-family: Trebuchet MS;"> Menstrual Cycle</h2> | ||
| − | <p style="font-family: Verdana;"> The monthly menstrual cycle of a womens' uterus consists of the menstrual, proliferative, and secretory phases. In menstruation, the womens' uterine lining sheds as it removes the egg from the body. Then during the proliferative phase, the surplus of estrogen stimulates the re-growth of the edometrium, also know as the uterine lining. And lastly, during the secretory phase, the levels of estrogen drop some while progesterone becomes the dominant hormone allowing the endometrium to be susceptible to pregnancy. Overall, this cycle lasts a total of 28 days and is large classified by the peaks and drops of estrogen and progesterone. </p> | + | |
| − | + | <p style="font-family: Verdana; float:left;"> The monthly menstrual cycle of a womens' uterus consists of the menstrual, proliferative, and secretory phases. In menstruation, the womens' uterine lining sheds as it removes the egg from the body. Then during the proliferative phase, the surplus of estrogen stimulates the re-growth of the edometrium, also know as the uterine lining. And lastly, during the secretory phase, the levels of estrogen drop some while progesterone becomes the dominant hormone allowing the endometrium to be susceptible to pregnancy. Overall, this cycle lasts a total of 28 days and is large classified by the peaks and drops of estrogen and progesterone. </p> | |
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<img src= "https://static.igem.org/mediawiki/2016/a/a7/T--MIT--KHB1177hormone-response.jpeg" alt = 'Hormone response diagram' style="width:250px;height:267px; float:left;" margin: 0 1.5%; class="rotate90"> | <img src= "https://static.igem.org/mediawiki/2016/a/a7/T--MIT--KHB1177hormone-response.jpeg" alt = 'Hormone response diagram' style="width:250px;height:267px; float:left;" margin: 0 1.5%; class="rotate90"> | ||
<h2 style="text-decoration:underline; font-family: Trebuchet MS;"> Estrogen</h2> | <h2 style="text-decoration:underline; font-family: Trebuchet MS;"> Estrogen</h2> | ||
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| − | <h1 style="background-color:#7ECEFD; -moz-border-radius: 15px; -webkit-border-radius: 15px; padding:15px; text-align: center; font-family: Trebuchet MS"> How | + | <br></br> |
| + | <h1 style="background-color:#7ECEFD; -moz-border-radius: 15px; -webkit-border-radius: 15px; padding:15px; text-align: center; font-family: Trebuchet MS"> How can our circuit detect hormones?</h1> | ||
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| + | <img src= "https://static.igem.org/mediawiki/2016/c/c6/T--MIT--KHB1177tretoere.jpeg" alt = 'TRE to pERE promoters' style="width:250px;height:267px; float:left;" margin: 0 1.5%; class="rotate90"> | ||
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| + | <p style="font-family:Verdana;"> | ||
| + | An important aspect of synthetic biology is having inducible systems so that the output is not produced constitutively. Since progesterone is a key biomarker of endometriosis and also one of the two components of the menstural cycle. We wanted to use the sensing of progesterone as a way to inhibit our system. In contrast we wanted to use the sensing of estrogen to activate our system. Currently, there had been some research on hormone inducible promoters, but this is largely lacking in the field of synthetic biology. We decided to tackle this problem by developing our own synthetic promoter, which was based off of the key components of the commmonly used synthetic promoter, Tetracyclin Response Element promoter (TRE). We kept the basic promoter elements, but rather than having tetO responsive sites, we used progesterone and estrogen responsive elements (PRE's and ERE's respectively. | ||
| + | </p> | ||
| + | <p style="font-family:Verdana;"><small><i>Read more about our design desicions for our inducible promoters: pERE, pPRE, and pHybrid.</i></small></p> | ||
| + | <br></br> | ||
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| + | <h1 style="background-color:#7ECEFD; -moz-border-radius: 15px; -webkit-border-radius: 15px; padding:15px; text-align: center; font-family: Trebuchet MS"> Do our synthetic promoters work?</h1> | ||
| + | <p style = "font-family:Verdana;"> SOme text goes here ya know. | ||
| + | </p> | ||
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| + | <p style="font-family:Verdana;"><small><i>Read more about our experiments testing the functionality of our promoters.</i></small></p> | ||
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| + | <h2 style="text-decoration:underline; font-family: Trebuchet MS;"> <center>MCF7 Induction of pEREx3 - eYFP</center></h2> | ||
| + | <img src= "https://static.igem.org/mediawiki/2016/4/40/T--MIT--KHB1177flourescence-fakeimg.jpeg" alt = 'TRE to pERE promoters' style="width:250px;height:267px; float:left;" margin: 0 1.5%; class="rotate90"> | ||
| + | <h2 style="text-decoration:underline; font-family: Trebuchet MS;"> <center>tHESC Induction of pEREx3 - eYFP</center></h2> | ||
| + | <img src= "https://static.igem.org/mediawiki/2016/4/40/T--MIT--KHB1177flourescence-fakeimg.jpeg" alt = 'TRE to pERE promoters' style="width:250px;height:267px; float:left;" margin: 0 1.5%; class="rotate90"> | ||
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<h2 style="text-decoration:underline; font-family: Trebuchet MS;"> Set Up </h2> | <h2 style="text-decoration:underline; font-family: Trebuchet MS;"> Set Up </h2> | ||
<p style="font-family: Verdana;"> Write about the experimental set up here </p> | <p style="font-family: Verdana;"> Write about the experimental set up here </p> | ||
Revision as of 03:02, 7 October 2016
How does endometriosis respond to hormones?
Estrogen
There are endogenous estrogen receptors in two forms: ER-alpha and ER-beta. When a healthy cell senses estrogen, the ER-alpha receptor is activated and triggers downstream responses by binding to different sites, such as an estrogen responsive element. Diseased cells respond in the same fashion as healthy cells.
Progesterone
There are also endoegenous progesterone receptors in two forms: PR-A and PR-B. When a healthy cell senses progesterone, its PR receptors are activated and trigger downstream responses by binding to different sites, such as a progesterone responsive element. However, in a diseased cell, while progesterone is present, it does not co-activate the progesterone receptors, and in turn does not result in any downstream effects.
How can our circuit detect hormones?
An important aspect of synthetic biology is having inducible systems so that the output is not produced constitutively. Since progesterone is a key biomarker of endometriosis and also one of the two components of the menstural cycle. We wanted to use the sensing of progesterone as a way to inhibit our system. In contrast we wanted to use the sensing of estrogen to activate our system. Currently, there had been some research on hormone inducible promoters, but this is largely lacking in the field of synthetic biology. We decided to tackle this problem by developing our own synthetic promoter, which was based off of the key components of the commmonly used synthetic promoter, Tetracyclin Response Element promoter (TRE). We kept the basic promoter elements, but rather than having tetO responsive sites, we used progesterone and estrogen responsive elements (PRE's and ERE's respectively.
Read more about our design desicions for our inducible promoters: pERE, pPRE, and pHybrid.
Do our synthetic promoters work?
SOme text goes here ya know.
Read more about our experiments testing the functionality of our promoters.
MCF7 Induction of pEREx3 - eYFP
tHESC Induction of pEREx3 - eYFP
Set Up
Write about the experimental set up here
Results
Write about the results here
