Difference between revisions of "Team:Linkoping Sweden/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 the earth's temperature. Our modern industrial civilization has increased the 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). 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's 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).

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?

Solution

Our project has made a huge progress during this year, we have designed a unique DNA construct and even started to work on the expression of the DNA construct in 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 Gibson assembly containing; a 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 CRISPR/Cas9-system which is induced by the LIP promoter. When the starch production is cut off, the fatty acid synthesis is naturally favored.

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.

The algae are first grown in pools covered by films (1) 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 UV light while streaming through transparent pipes (2).

The algae, with the activated CRISPR/Cas9 within them, end up in different pools (3) where the whelming films are no longer needed.

The oil extracted out of the algae (4) is later turned into biodiesel (5) and is capable to provide our polluted 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.

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. 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 (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.

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.