Difference between revisions of "Team:Pitt/Practices"

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<h2> Human Practices </h2>
 
<h2> Human Practices </h2>
  
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<div class="click"><h3>Implementation</h3>
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<p>When considering the many implications that releasing a paper-based sensor as a commercial product might have, the Pitt iGEM team selected two areas of focus. First and foremost, our goal was to create a very cheap sensor that is easily transportable. To this end, we designed our own protocol for generating such sensors, rather than purchasing expensive commercial extracts. With our protocol, we were able to create extracts in as little as three days in large quantity. With some simple automation of the process, we estimate that 100 paper strips can be created for less than a dollar, not including instrument time costs. With such cheap production available, this makes the final product affordable for many, especially considering the simple transport of the sensors.</p>
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</p>The second hurdle we attempted to overcome with the design of our project was the possibility of false positives. With many cell-based assays, it is difficult to have a binary response, as a gradient of concentrations will produce a gradient of responses. We attempted to differentiate between a positive and negative response by building a amplification and quenching circuit, where the amplification would ensure a positive result would be visible, and the quencher would make negative results indistinguishable from no signal. The amplification loop consists of a T3 promoter driving the production of T3 RNA Polymerase. By having the sensing system output T3 RNA Polymerase, the amplification loop with create exponentially more of the polymerase, leading to a stronger signal from a reporter construct based on the T3 promoter. The quenching is based on simple competitive inhibition of T7 or T3 RNA polymerase by a DNA "dumbell" hairpin decoy. By varying the amount of decoy in the reaction, varying amounts of basal transcription and translation can effectively be quenched. For more information on this part of the project, click <a href="2015.igem.org/Team:Pitt/amplification">here.</a></p>
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<p>Paper-based sensors capable of detecting diseases and pollutants will interact with society in a myriad of ways.  In order to facilitate the growth and utilization of our project, the 2015 Pitt iGEM team intends on predicting and accounting for the human factors that will influence the success of our paper-based sensors.</p>
 
<p>Paper-based sensors capable of detecting diseases and pollutants will interact with society in a myriad of ways.  In order to facilitate the growth and utilization of our project, the 2015 Pitt iGEM team intends on predicting and accounting for the human factors that will influence the success of our paper-based sensors.</p>
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<p>Clearly, there are a variety of interactions that paper-based sensors will have with the surrounding human environment; there are likely some outcomes that we cannot predict—the market inherently gives products their most thorough analysis—but we have certainly found the most profound human factors that will affect the way our product interacts with the surrounding society.  Our identification and analysis of these factors combined with our complementary product design will allow for a sustainable and successful commercial launch of paper-based sensors.</p> </div>
 
<p>Clearly, there are a variety of interactions that paper-based sensors will have with the surrounding human environment; there are likely some outcomes that we cannot predict—the market inherently gives products their most thorough analysis—but we have certainly found the most profound human factors that will affect the way our product interacts with the surrounding society.  Our identification and analysis of these factors combined with our complementary product design will allow for a sustainable and successful commercial launch of paper-based sensors.</p> </div>
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<div class="color"><h3>Implementation</h3>
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<p>When considering the many implications that releasing a paper-based sensor as a commercial product might have, the Pitt iGEM team selected two areas of focus. First and foremost, our goal was to create a very cheap sensor that is easily transportable. To this end, we designed our own protocol for generating such sensors, rather than purchasing expensive commercial extracts. With our protocol, we were able to create extracts in as little as three days in large quantity. With some simple automation of the process, we estimate that 100 paper strips can be created for less than a dollar, not including instrument time costs. With such cheap production available, this makes the final product affordable for many, especially considering the simple transport of the sensors.</p>
 +
 +
</p>The second hurdle we attempted to overcome with the design of our project was the possibility of false positives. With many cell-based assays, it is difficult to have a binary response, as a gradient of concentrations will produce a gradient of responses. We attempted to differentiate between a positive and negative response by building a amplification and quenching circuit, where the amplification would ensure a positive result would be visible, and the quencher would make negative results indistinguishable from no signal. The amplification loop consists of a T3 promoter driving the production of T3 RNA Polymerase. By having the sensing system output T3 RNA Polymerase, the amplification loop with create exponentially more of the polymerase, leading to a stronger signal from a reporter construct based on the T3 promoter. The quenching is based on simple competitive inhibition of T7 or T3 RNA polymerase by a DNA "dumbell" hairpin decoy. By varying the amount of decoy in the reaction, varying amounts of basal transcription and translation can effectively be quenched. For more information on this part of the project, click <a href="2015.igem.org/Team:Pitt/Amplification/Project">here.</a></p>
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Revision as of 08:20, 18 September 2015

Human Practices

Considerations

Paper-based sensors capable of detecting diseases and pollutants will interact with society in a myriad of ways. In order to facilitate the growth and utilization of our project, the 2015 Pitt iGEM team intends on predicting and accounting for the human factors that will influence the success of our paper-based sensors.

One observer considering the ethics of at-home disease tests might ask us, “is it ethical to produce a test that could give a consumer a false positive indication of cancer or illness? Such a result could lead to significant anxiety, grief or depression—potentially damaging to the consumer—until the issue is resolved by a medical practitioner.” This concern is certainly an issue for our paper-based sensors; the issue would primarily be resolved with clear and obvious disclaimers that inform our consumer that the tests are not 100% accurate and that final diagnoses should always be administered by certified doctors. Furthermore, the tests are so cheap to produce that they could be affordably packaged two or three at a time to provide multiple tests which would decrease the chances of a false positive exponentially. However, there is a complementary question of ethics that we must pose in favor of our products: “is it ethical to value some peoples’ state of mind over the potential to save others’ lives or improve their prognoses?” The answer to this question—when posed to sane humans—is overwhelmingly ‘no’. Paper-based sensors provide an opportunity to help members of society in a significant and dynamic way, and the potential for false positives cannot hold back such an advancement.

Another important human factor aside from ethics is the safety of the paper-based sensors. Skeptics would be quick to point out any flaw in our product’s safety, leading to fewer customers at best—and public backlash at worst. However, our product was designed with safety in mind; the tests for blood, urine and water work by placing a sample from the source onto the paper test. This ensures that the tests never make direct contact with the patient or water source, preventing any potential backwards contamination. Therefore, the only potential safety issue associated with the paper-based sensors would be the extraction of a blood sample, and that process has been streamlined over decades to ensure a safe process. The conclusion based on our safety analysis is that paper-based sensors provide a safe and non-invasive method of disease and pollution detection.

Since it has been verified that the potential product is both safe and ethical, we must determine whether our product is economically feasible for commercial production. An important question that an investor would likely pose to our team might be, “is your project scalable for mass-production?” The answer is yes; many of the steps in the production of the paper-based sensors will be streamlined into continuous processes. These continuous processes include steady-state bioreactors culturing the proper bacteria strain, as well as the distribution of the cell-free systems to paper through mechanical pipettes and the subsequent flash freezing of the paper-based sensors with liquid nitrogen. The systematic verification of product quality will also be performed continuously. The remaining processes will be easily carried out in large batch form; the main batch processes include centrifuging and lysing the bacterial cultures and freeze drying the paper-based sensors. Although these large scale processes will ensure a high throughput—and therefore a significant economy of scale—the important factor which will determine our magnitude of success would be the market price of the tests. It has been shown by researchers that cell-free systems freeze dried on paper can be produced for less than one dollar per reaction. This ensures that a market price of less than ten dollars per test is feasible. This is an easily affordable test, which will help to ensure access to our product regardless of socioeconomic status.

The economic feasibility of our project is not only favorable for an entrepreneurial entrance into the healthcare market, but it is also conducive to social justice. Currently, low socioeconomic status prevents some people from seeking necessary healthcare—especially if they are unsure if they are afflicted with a given suspected disease. Since our paper-based sensors can be sold at a low market price, they will be available to be purchased by anyone. This availability ensures that most people are able to determine whether they are afflicted with their suspected ailment.

Clearly, there are a variety of interactions that paper-based sensors will have with the surrounding human environment; there are likely some outcomes that we cannot predict—the market inherently gives products their most thorough analysis—but we have certainly found the most profound human factors that will affect the way our product interacts with the surrounding society. Our identification and analysis of these factors combined with our complementary product design will allow for a sustainable and successful commercial launch of paper-based sensors.

Implementation

When considering the many implications that releasing a paper-based sensor as a commercial product might have, the Pitt iGEM team selected two areas of focus. First and foremost, our goal was to create a very cheap sensor that is easily transportable. To this end, we designed our own protocol for generating such sensors, rather than purchasing expensive commercial extracts. With our protocol, we were able to create extracts in as little as three days in large quantity. With some simple automation of the process, we estimate that 100 paper strips can be created for less than a dollar, not including instrument time costs. With such cheap production available, this makes the final product affordable for many, especially considering the simple transport of the sensors.

The second hurdle we attempted to overcome with the design of our project was the possibility of false positives. With many cell-based assays, it is difficult to have a binary response, as a gradient of concentrations will produce a gradient of responses. We attempted to differentiate between a positive and negative response by building a amplification and quenching circuit, where the amplification would ensure a positive result would be visible, and the quencher would make negative results indistinguishable from no signal. The amplification loop consists of a T3 promoter driving the production of T3 RNA Polymerase. By having the sensing system output T3 RNA Polymerase, the amplification loop with create exponentially more of the polymerase, leading to a stronger signal from a reporter construct based on the T3 promoter. The quenching is based on simple competitive inhibition of T7 or T3 RNA polymerase by a DNA "dumbell" hairpin decoy. By varying the amount of decoy in the reaction, varying amounts of basal transcription and translation can effectively be quenched. For more information on this part of the project, click here.