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Designing a bacterial nitric oxide sensor to detect Leishmania parasites inside macrophages and eliminate them

$190
Raised of $2,345 Goal
9%
Ended on 8/13/19
Campaign Ended
  • $190
    pledged
  • 9%
    funded
  • Finished
    on 8/13/19

About This Project

Leishmaniasis is an intracellular parasitic disease caused by Leishmania, that causes up to one million new cases annually and has an annual mortality rate of 65000 people. We hypothesize that we can genetically engineer a bacterium to detect the presence of Leishmania-infected macrophages using a nitric oxide sensor, and in response, express iron chelating agents (to compete with Leishmania for iron) and consequently eliminate them from the affected cells using hydrolytic enzymes.


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What is the context of this research?

The inspiration behind the project is to engineer a therapeutic bacteria that can control and monitor the various biochemical processes that are going on in our body and thus effectively limit the growth of pathogens and cancer cells in the body without harming the host. We have specifically decided to target the dreadful disease leishmaniasis, known locally as kala azar. It is the second largest parasitic killer in the world and is highly endemic to the impoverished regions of Bengal and Bihar in India. To target this disease, we aim to use the inflammatory biomarker, Nitric oxide (NO), which is downregulated inside Leishmania-infected macrophages. We hypothesise that we can use this characteristic NO level as a signal to detect Leishmania-infected macrophages inside the body.

What is the significance of this project?

Despite a concerted effort by the international community to weed out this life altering disease, Leishmania continues to rampage in almost 47 countries endangering approx 200 million people worldwide. A few recent examples are presented here as links: Kenya(23/4/19); Sweden(4/5/19); Pakistan(6/2/19). In India itself,1 in 10 people are at risk of contracting the disease. Many treatment options are available but each of them havemajor disadvantages, like amphotericin B (expensive, antibiotic resistance) and sodium stibogluconate (serious side effects). We aim to develop a cheap and widely accessible diagnostic and therapeutic tool using (GM) E.coli bacteria to target Leishmania-infected macrophages without causing major side effects or giving rise to resistance development.

What are the goals of the project?

Our project's hypothesis is that Leishmania-infected macrophages can be detected by engineering a bacterium with a nitric oxide (NO) sensor which is activated only when intracellular NO level falls within a certain concentration range. This NO level range is unique to Leishmania-infected macrophages and ensures that our targeted response is specific to these cells only. This sensor activates the expression of an iron chelator (to reduce iron available for leishmania) and a hydrogen peroxide-producing enzyme to kill both the Leishmania and the engineered bacteria within the macrophage. To test this hypothesis, we shall treat Leishmania-infected macrophages with our engineered therapeutic bacteria to check for Leishmania clearing inside macrophages. Click on this link for more details.

Budget

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The budget is mainly comprised of 3 components-

1) Lab Reagents and Enzymes- These include bacterial culture media, antibiotics, fetal bovine serum and DMEM (for macrophage and leishmania culture), DETA/NO (Nitric oxide donor), Griess Reagent and restriction enzymes.

2) Kits- We need to use "Intracellular Nitric Oxide Assay Kit" (to measure intracellular NO levels) and "Gel Extraction Kit" for purification of DNA from gels.

3) Giant Jamboree- The Giant Jamboree is the ultimate event of the iGEM competition, held in MIT, Boston, USA. This includes the poster presentation of our project. The Jamboree is attended by every participating iGEM team and it is mandatory for each team to send at least one team member and hence we have included the registration fee for one team member.

Endorsed by

It's always a ground breaking in research when a group of young scholars draws "hypothesis", jumps unbound to"test" it in lieu of convention of proving a theory.By this test of hypothesis on Liesmaniasis, iGEM 2019 group has placed itself in a very high confidence level as example. Being with them from beginning in lab,induced by their overwhelming enthusiasm & total involvement, I’m confident they will reach their goal & try hard to give back a Kala Azar free community. Hence, I feel to support them in all possible ways.
Being a member of this project, I am really feeling very excited. Students are doing very well to achieve the goal. Wishing them all the best for future endeavors.
I think this project embodies a very innovative way to address a crucial public health challenge. Using synthetic biology to find new ways to treat leishmaniasis is an exciting plan. The IISER Kolkata iGEM team consists of the right mix of enthusiasm and expertise to get this work done. I endorse the efforts of the team and wish them an exciting project ahead.
I am really exited about this very innovative yet challenging project. I believe it will answer very important question in the field of Leishmania Biology. I fully endorse the capability and the required knowledge that this group of iGEM students in solving this. I wish them very best of luck in achieving their goals.

Flag iconProject Timeline

Currently we are training for the various experiments and have started ordering laboratory reagents. Our project will have 3 phases. Phase I (Mid June-Mid July) will include genetic construct assembly, bacterial transformation and testing the genetic construct functionality using GFP. Phase II (Mid July-end of August) will focus on the determination of intracellular NO levels in uninfected and infected macrophages. Phase III will focus on aerobactin production and bacterial treatment efficiency.

Jun 10, 2019

Project Launch

Jun 24, 2019

Gene Sequencing to check engineered genetic constructs after biobrick assembly

Jul 08, 2019

Obtaining bacteria transformed with engineered genetic constructs

Jul 14, 2019

Project Launched

Jul 15, 2019

Testing Functionality of genetic constructs using GFP fluorescence

Meet the Team

Priyanuj Bordoloi
Priyanuj Bordoloi
Undergraduate

Affiliates

Indian Institute of Science Education and Research, Kolkata
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Diptomit Biswas
Diptomit Biswas
Undergraduate

Affiliates

Indian Institute of Science Education and Research, Kolkata
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Abhirami A G
Abhirami A G
Undergraduate

Affiliates

Indian Institute of Science Education and Research, Kolkata
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Atharva Paranjape
Atharva Paranjape
Undergraduate

Affiliates

Indian Institute of Science Education and Research, Kolkata
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Ipshita Gayen
Ipshita Gayen
Undergraduate

Affiliates

Indian Institute of Science Education and Research, Kolkata
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Sumit Garai
Sumit Garai
Undergraduate

Affiliates

Indian Institute of Science Education and Research, Kolkata
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Roohani B
Roohani B
Undergraduate

Affiliates

Indian Institute of Science Education and Research, Kolkata
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Aniruddh Deshpande
Aniruddh Deshpande
Undergraduate

Affiliates

Indian Institute of Science Education and Research, Kolkata
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Angel Mary C.T
Angel Mary C.T
Undergraduate

Affiliates

Indian Institute of Science Education and Research, Kolkata
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Pinaki Chanda
Pinaki Chanda
Undergraduate

Affiliates

Indian Institute of Science Education and Research, Kolkata
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Urvashi Chitnavis
Urvashi Chitnavis
Undergraduate

Affiliates

Indian Institute of Science Education and Research, Kolkata
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Putchala Ravi Kiran
Putchala Ravi Kiran
Undergraduate

Affiliates

Indian Institute of Science Education and Research, Kolkata
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Team Bio

We are a team of 12 undergraduates from Indian Institute of Science Education and Research (IISER) Kolkata. It was established by the MHRD in 2006 and is an Institute of National Importance. It is one of seven IISERs set up by the Govt. of India. Our team consists of students from various fields (STEM); this enables us to garner a deeper understanding while looking at the problem from different perspectives. We are driven by one single purpose- to use synthetic biology as a harbinger of change.

Priyanuj Bordoloi

I love the interdisciplinary aspect of iGEM, it's so much more than a "synthetic biology" project. Even though Biology evolves by the means of random tinkering, yet it always boils down to a defined set of circuit elements that obey simple general design principles. And there is a certain beauty in seeing such systems in mathematical terms. iGEM provides me with the opportunity to pursue this interest. Also, I get to do cool bio stuff which I only get to see in movies :p ⁣

Diptomit Biswas

My vision of the future of therapeutic science entails the use of artificial cellular machines to control and monitor on a nanoscale level the various biochemical processes that are going on in our body. These nanomachines would be highly effective in limiting the growth of pathogens and cancer cells inside our body without harming the host itself and that's why I want to take part in iGEM. It provides the perfect platform to test and share such wonderful ideas with the rest of the scientific community⁣⁣.⁣⁣

Abhirami A G

iGEM demands an interdisciplinary approach to social issues, which I always wanted to pursue. When principles of engineering are brought to biology, it meets its completion. Personally, participating in iGEM is one of the best things that can happen to me.⁣

Atharva Paranjape

As an undergraduate researcher, I am constantly looking for learning opportunities. I love to try out new topics and immerse myself in them. Personally, I have a special attraction to biology and physics. iGEM is the perfect interdisciplinary playground for me and I intend to make the most of it, for myself and my team⁣

Ipshita Gayen

I see myself as a hardcore biologist with a massive inclination towards the field of Ecology. Nevertheless, my curiosity likes to dive deep into other fields of biology too, and iGEM, with its roots in synthetic biology, provides me with the right opportunity. It's a platform that provides me with a plethora of topics to explore, ranging from genetics to mathematical modelling (which I hope to learn from my juniors). Moreover, it's a wonderful chance to enrich lab skills, interact with people from various disciplines and share diverse perspectives. Last but not least, I want to contribute my small part in the betterment of society, which is what iGEM strives to achieve; encouraging young minds to tackle and find solutions to the small and large problems faced in everyday life. ⁣⁣

Sumit Garai

Well, the coolest thing about iGEM is the freedom that we get. We decide our own topic to work on, we decide our own lab setup, and instead of professors telling us what to do, we tell them what we do😂⁣⁣

Roohani B

It's really exciting to compete to have the coolest biotechnology project against teams from around the world. This is a great opportunity to gain skills and explore a lot of different areas in the hopes of identifying a field that interests me and realize that real research is far different from the lab courses we take in college. Taking part in such a project shall not only kindle an interest in research but also help me think in a way biologists should.

Aniruddh Deshpande

I am fascinated by a wide spectrum of topics other than my primary interest itself i.e., Physics. I am intrigued by all the STEM fields.⁣⁣This is where iGEM gives me an opportunity to apply my knowledge in physics, maths, and coding to biological systems which is amazing! I get to do interdisciplinary work at an early stage in my undergrad course.

Angel Mary C.T

Biology, as a subject inspires me. When I consider the bigger picture, I see this subject as one that can help me to create a direct impact on society. Ultimately, I want to do good to the society. When I got the mail that I
got selected for iGEM, l knew that this could be a beginning.⁣

Pinaki Chanda

iGEM is a good platform for students like us to present our idea in front of the world. Moreover, I’m excited to learn new things and work with the team. I hope the journey of the iGEM will be one of the most wonderful experiences. As iGEM is a competition, our primary goal for the next few months is to secure a good position there but parallel to that our team has a strong determination to “Heal the world and make it a better place”.⁣

Urvashi Chitnavis

It's fascinating, the extent to which life can be manipulated today - and it's only going to get better. I read about all these amazing advances in science, and get so frustrated that I'm not yet a part of them. iGEM is the opportunity of a lifetime, a chance to finally contribute a tiny bit to the science I love so much and learn an immeasurable amount in the process.

Putchala Ravi Kiran

Among many planets known till date, Earth is the only planet which supports life. Each and every living organism, from single-celled bacteria to complex mammalian systems, has a diversified genome which makes them unique. The regulatory pathway for such gene expression producing a variety of proteins makes this field extremely interesting to study and play.⁣iGEM gives me a platform for studying different regulatory processes and play with different combinations and mutated gene sequences.⁣

Additional Information

Relevant details about the project can be found here on our website. For clarification on any aspect of the project, please reach us here at team.igem@iiserkol.ac.in . We would we happy to address your questions to the best of our efforts.


Project Backers

  • 8Backers
  • 9%Funded
  • $190Total Donations
  • $23.75Average Donation
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