Difference between revisions of "Team:Linkoping Sweden/Description"

Latest revision as of 20:41, 6 December 2016

Project Description

Overview

When conducting a project there are many aspects to consider. LiU iGEM has been working within different areas such as human practices, ethics and experimental design to obtain the best possible results. With human practices the team has reached out to the public by attending different exhibitions, given information meetings and presentations. Furthermore, we have actively used social media to reach out to a broader audience by frequently updating about our progress. Apart from informing the public, we have been in contact with both the Technical Research Institution of Sweden and The Swedish Gene Technology Advisory Board. The purpose was to inform them about our project and to get feedback. With this in mind a pamphlet containing environmental, economic and health values was put together. The safety level of our project was confirmed to be at the lowest risk by the Swedish Work Environmental Authority.

Collaborations are of great importance according to LiU iGEM, not only with authorities but also with other iGEM teams. We have collaborated with other teams by sharing our knowledge about ethics and safety regarding the usage of CRISPR/Cas9 in Chlamydomonas reinhardtii and what needs to be considered in sustainable large-scale productions. Exchange of different protocols, improvement of data and ideas to make goals possible are subjects included in our collaborations.

Problem

The International Panel of Climate Change reported conclusions that human activities over the past 50 years have led to the increase of greenhouse gases such as carbon dioxide, nitrogen oxide and methane (1) resulting in augmentation of the earth's temperature.

Algae biofuels have proven to be a possible substitute for fossil fuels. When utilizing algae there are some obstacles to overcome, for instance improvement and identification of suitable strains as in terms of oil productivity and to insure isolation from other crops (3).

There are plenty of questions concerning this area, and the main focus will be the following:

How will an increased lipid synthesis be attained without damaging the reproductivity of the algae?
How could the biodiesel production from our algae be up scaled?

Solution

Our solution to the problem includes the design of a unique DNA construct and the expression of the DNA construct in Chlamydomonas reinhardtii. Before a large-scale production of the product is possible, there are certain hindrances that need to be overcome.

We have been working with the model algae C. reinhardtii into which we have transformed a DNA construct (see figure 1), through Gibson assembly containing; a light inducible promoter LIP, a Cas9 sequence and a sgRNA sequence. These sequences combined result in a knock out of a gene coding for starch synthesis, through the CRISPR/Cas9-system which is induced by the LIP promoter. When the starch production is cut off, the fatty acid synthesis is naturally favored.

Figure 1. Transformation of DNA construct into C. reinhardtii followed by light inducement and optimization by CRISPR/Cas9. Resulting in a magnified C. reinhardtii.

The idea with the project is to enable an economically sustainable production of biodiesel from algae. The production itself is easily described with a cycle of events that we propose. The final event, when the algae consume carbon dioxide from the environment, the production circle is complete, seen below in Figure 2.

Figure 2. The cycle of biodiesel production from algae.

The algae are first grown in pools covered by films (step 1 in figure 2) removing high intensity light, preventing the initiation of the CRISPR/Cas9-system. This phase enables the algae to grow to a proper size before being exposed to ultra violet light while streaming through transparent pipes (step 2). The whelming films are removed resulting in an activation of the CRISPR/Cas9 system (step 3). The oil extracted from the algae ('step 4) is later turned into biodiesel (step 5) and is capable to provide our polluted planet with a green alternative for both individuals and the industry (step 7-1). The biodiesel production we could offer would be a helping hand in the long-term substitution of fossil fuels.

Even if the oil extraction (step 4) and the biodiesel production (step 5) techniques are constantly improved there are often residues of protein and glycerin to some extent. Protein residues are mainly used as feeding of fish farms and other animals in the agricultural industry. The residues enrich the food with protein and could also be used as a protein supplement for humans. The glycerin products separated during the biodiesel production (step 5) have a substantial area of use for example in the food industry where the glycerin serves as a sweetener, stabilizer and humectant among others, as well as the pharmaceutical industry when manufacturing drugs with improving smoothness, with humectant and with providing lubrication. New areas of application have recently been found, for instance glycerin has been proven to suppress inflammatory responses in the body.

Assuming that our DNA construct would be expressed successfully the CRISPR-biofuel would not be far away. Large scale production of the biodiesel from C. reinhardtii could be the answer to a sustainable environment in the future.

References

1. NASA - Global Climate Change. A blanket around the Earth. Retrieved 1 Sep. 2016 from: http://climate.nasa.gov/causes/

2. Michael Hannon, Javier Gimpel, Miller Tran, Beth Rasala, and Stephen Mayfield (2010). Biofuels from algae: challenges and potential. US National Library of Medicine National Institutes of Health. PMCID: PMC3152439 http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3152439/

3. Myung, et. al., Effects of dietary substitution of fishmeal with the combined dry microalgae, Nannochloropsis oceanica (NO) biomass residue and casein on growth and body composition of juvenile abalone (Haliotis discus)., 2014 http://www.feednavigator.com/R-D/Residue-of-microalgae-derived-biofuel-could-work-as-fishmeal-alternative-in-aquafeed

4. Fischer Günther and Prieler Syler, Agricultural by-products assicoated with biofuel production chains (report D5.1), 2009

5. The Soap and Detergent Association, Glycerine: an overwiev, http://www.aciscience.org/docs/glycerine_-_an_overview.pdf

6. thechemicalcompany, Global Bonds in Chemistry, Glycerine, 2016, https://thechemco.com/chemical/glycerine/

LiU iGEM is proudly sponsored by

@@ Line 1: / Line 1: @@
 {{Linkoping_Sweden}}
-<div class="column full_size">
-===Overview===
+==Project Description==
 ----
-When conducting a project there are many aspects to consider. LiU iGEM has been working within different areas such as human practice, ethics and experimental design for best possible results.
-With human practice the team has reached out to the public by attending different exhibitions, offered information meetings and presentations. Furthermore, we have actively used social media to reach out to a broader audience. Apart from informing the public, we have been in contact with both Sweden’s Technical Research Institution and Genetics Board. This was to inform them about our project and get feedback, from this we constructed a pamphlet containing environmental, economic and health values. With help from the Swedish Work Environmental Authority the safety level of our project was confirmed to be at the lowest risk.
-LiU iGEM find a great importance in collaborations, not only with authorities but also with other iGEM Teams. We have collaborated with other teams by sharing our knowledge of ethics and safety regarding the usage of CRISPR/Cas9 in ''Chlamydomonas reinhardtii'' and what to consider in a large-scale production and sustainability. Collaborations have also included exchange of different protocols, improvement of data and ideas to make their goals possible.
-Another ambition has been to contribute to iGEM BioBrick Database by designing three new BioBricks according to standard model. Also, we had the ambition to improve a Cas9 BioBrick by creating a construct with a related promotor.
+===Overview===
+When conducting a project there are many aspects to consider. LiU iGEM has been working within different areas such as human practices, ethics and experimental design to obtain the best possible results.
+With human practices the team has reached out to the public by attending different exhibitions, given information meetings and presentations. Furthermore, we have actively used social media to reach out to a broader audience by frequently updating about our progress. Apart from informing the public, we have been in contact with both the Technical Research Institution of Sweden and The Swedish Gene Technology Advisory Board. The purpose was to inform them about our project and to get feedback. With this in mind a pamphlet containing environmental, economic and health values was put together. The safety level of our project was confirmed to be at the lowest risk by the Swedish Work Environmental Authority.
+Collaborations are of great importance according to LiU iGEM, not only with authorities but also with other iGEM teams. We have collaborated with other teams by sharing our knowledge about ethics and safety regarding the usage of CRISPR/Cas9 in ''Chlamydomonas reinhardtii'' and what needs to be considered in sustainable large-scale productions. Exchange of different protocols, improvement of data and ideas to make goals possible are subjects included in our collaborations.
 ===Problem===
-<p>The International Panel on Climate Change reported conclusions that human activities over the past 50 years has led to the increase in the earth’s temperature. Our modern industrial civilization has increased CO2-levels from 280 ppm to 400 ppm, much of the harm has been a result of human-produced greenhouse gases such as CO2, N2O and Methane (1).
+The International Panel of Climate Change reported conclusions that human activities over the past 50 years have led to the increase of greenhouse gases such as carbon dioxide, nitrogen oxide and methane (1) resulting in augmentation of the earth's temperature.
-For example, any carbon based fuel that is burned will convert carbon to CO2, if not properly stored it will be released into the atmosphere thereby increasing CO2 levels. The earth surface is emitting IR-radiation and the CO2 prevent it from leaving the Earth surface and released into space by absorbing the radiation/heat, consequently contributing to an increase in temperature (2).</p>
+Algae biofuels have proven to be a possible substitute for fossil fuels. When utilizing algae there are some obstacles to overcome, for instance improvement and identification of suitable strains as in terms of oil productivity and to insure isolation from other crops (3).
+There are plenty of questions concerning this area, and the main focus will be the following:
+* How will an increased lipid synthesis be attained without damaging the reproductivity of the algae?
+* How could the biodiesel production from our algae be up scaled?
+===Solution===
+Our solution to the problem includes the design of a unique [[Team:Linkoping_Sweden/Design|DNA construct]] and the expression of the DNA construct in ''[[Team:Linkoping_Sweden/Description/Chlamydomonas_reinhardtii|Chlamydomonas reinhardtii]]''. Before a large-scale production of the product is possible, there are certain hindrances that need to be overcome.
+We have been working with the model algae ''C. reinhardtii'' into which we have transformed a DNA construct (see figure 1), through [[Team:Linkoping_Sweden/Description/Gibson_assembly|Gibson assembly]] containing; a [[Team:Linkoping_Sweden/Design#num2|light inducible promoter]] LIP, a Cas9 sequence and a sgRNA sequence. These sequences combined result in a knock out of a gene coding for starch synthesis, through the [[Team:Linkoping_Sweden/Description/CRISPR/Cas9-system|CRISPR/Cas9-system]] which is induced by the LIP promoter. When the starch production is cut off, the fatty acid synthesis is naturally favored.
-There has been lots of discussions about other alternatives for fossil fuels and algae biofuels has proven to be a possible alternative. By utilizing algae there are some difficulties to overcome, for instance improvement and identification of strains - in terms of oil productivity and protection of crops (3).
-There are plenty of questions concerning this area, but the focus will be on answering these questions: How will an increased lipid synthesis be attained without damaging the reproductivity? How will this be controlled? Lastly, how will this be used in a larger scale?
 [[File:T--Linkoping Sweden--linjart.png|700 px|center]]
-'''Figure 1.''' Transformation of DNA construct into ''C. reinhardtii'' followed by light inducement and optimization by CRISPR/Cas9. Resulting in a magnified ''C. reinhardtii''.
-===Solution===
+<center>'''Figure 1.''' Transformation of DNA construct into ''C. reinhardtii'' followed by light inducement and optimization by CRISPR/Cas9. Resulting in a magnified ''C. reinhardtii''.</center>
-Our project has made a huge progress during this year, we have designed a unique [[Team:Linkoping_Sweden/Design|DNA construct]] and even started to work on the expression of the DNA construct in [[Team:Linkoping_Sweden/Description/Chlamydomonas_reinhardtii|Chlamydomonas reinhardtii]]. There is still some progress that needs to be achieved before a large-scale production of the product is possible. The importance with a large-scale production is to consider the production line and related aspects in an early state. In this section the future biodiesel production from ''C. reinhardtii'' is described.
-We have been working with the model algae ''C. reinhardtii'' into which we have transformed a DNA construct, through [[Team:Linkoping_Sweden/Description/Gibson_assembly|Gibson assembly]] containing; a [[Team:Linkoping_Sweden/Design#num2|light inducible promoter]] called LIP, a Cas9 sequence and an sgRNA sequence. These sequences combined results in an inactivation of a gene coding for starch synthesis, through the [[Team:Linkoping_Sweden/Description/CRISPR/Cas9-system|CRISPR/Cas9-system]] which is induced by the LIP promoter. When the starch production is cut off, the fatty acid synthesis is naturally favored.
+The idea with the project is to enable an economically sustainable production of biodiesel from algae. The production itself is easily described with a cycle of events that we propose. The final event, when the algae consume carbon dioxide from the environment, the production circle is complete, seen below in Figure 2.
-Our project enables an economical sustainability in the production of biodiesel from algae. The production itself is easily described with a cycle of events. The final event, when the algae consume carbon dioxide from the environment, the production circle is complete, seen below.
 [[File:T--Linkoping Sweden--Kretslopp.png|900px|center]]
-'''Figure 2.''' The cycle of biodiesel production from algae.
+<center>'''Figure 2.''' The cycle of biodiesel production from algae.</center>
-The algae are first grown in pools covered by films (1) preventing the initiation of the CRISPR/Cas9-system. This phase enables the algae to grow to a proper size before being exposed to UV light while streaming through transparent pipes (2).
+The algae are first grown in pools covered by films (step 1 in figure 2) removing high intensity light, preventing the initiation of the CRISPR/Cas9-system. This phase enables the algae to grow to a proper size before being exposed to ultra violet light while streaming through transparent pipes (step 2). The whelming films are removed resulting in an activation of the CRISPR/Cas9 system (step 3). The oil extracted from the algae ('step 4) is later turned into biodiesel (step 5) and is capable to provide our polluted planet with a green alternative for both individuals and the industry (step 7-1). The biodiesel production we could offer would be a helping hand in the long-term substitution of fossil fuels.
+Even if the oil extraction (step 4) and the biodiesel production (step 5) techniques are constantly improved there are often residues of protein and glycerin to some extent. Protein residues are mainly used as feeding of fish farms  and other animals in the agricultural industry. The residues enrich the food with protein and could also be used as a protein supplement for humans. The glycerin products separated during the biodiesel production (step 5) have a substantial area of use for example in the food industry where the glycerin serves as a sweetener, stabilizer and humectant among others, as well as the pharmaceutical industry when manufacturing drugs with improving smoothness, with humectant and with providing lubrication. New areas of application have recently been found, for instance glycerin has been proven to suppress inflammatory responses in the body.
+Assuming that our DNA construct would be expressed successfully the CRISPR-biofuel would not be far away. Large scale production of the biodiesel from ''C. reinhardtii'' could be the answer to a sustainable environment in the future.
+===References===
+----
+. NASA - Global Climate Change. A blanket around the Earth. Retrieved 1 Sep. 2016 from: http://climate.nasa.gov/causes/
-The algae, with the activated CRISPR/Cas9 within them, end up in different pools (3) where the whelming films are no longer needed.
+. Michael Hannon, Javier Gimpel, Miller Tran, Beth Rasala, and Stephen Mayfield (2010). Biofuels from algae: challenges and potential. US National Library of Medicine National Institutes of Health. PMCID: PMC3152439
+http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3152439/
-The oil extracted out of the algae (4) is later turned into biodiesel (5) and is capable to provide our dying planet with a green alternative for both individuals and the industry (7-1). The biodiesel production we could offer would be a helping hand in the long-term substitution of fossil fuels.
+. Myung, et. al., Effects of dietary substitution of fishmeal with the combined dry microalgae, Nannochloropsis oceanica (NO) biomass residue and casein on growth and body composition of juvenile abalone (Haliotis discus)., 2014  http://www.feednavigator.com/R-D/Residue-of-microalgae-derived-biofuel-could-work-as-fishmeal-alternative-in-aquafeed
-Even if the oil extraction (4) and the biodiesel production (5) techniques are constantly improved there are often residues of protein and glycerin to some extent.
+. Fischer Günther and Prieler Syler, Agricultural by-products assicoated with biofuel production chains (report D5.1), 2009
-Protein residues are mainly used as feeding of fish farms  and other animals in the agricultural industry. The residues enrich the food with protein and could also be used as a protein supplement for humans.
+. The Soap and Detergent Association, Glycerine: an overwiev, http://www.aciscience.org/docs/glycerine_-_an_overview.pdf
-The glycerin products separated during the biodiesel production (5) have a huge area of use for example in the food industry where the glycerin serves as a sweetener, stabilizer and humectant among others, and also within the pharmaceutical industry when manufacturing drugs with improving smoothness, with humectant and with providing lubrication. New areas of application have recently been found, for instance glycerin has been proven to suppress inflammatory responses in the body.
+. thechemicalcompany, Global Bonds in Chemistry, Glycerine, 2016, https://thechemco.com/chemical/glycerine/
-The large-scale production of the biodiesel from C. reinhardtii may lie in the future, but if the DNA construct would be expressed as expected, the CRISPR-biofuel would not be far away.
 {{Linkoping_Sweden/Footer}}