2018: Developing a Mutational Library of and a Drop-Out Screen for Aβ-42
Our proposed project seeks to investigate amyloid-beta aggregopathy in Alzheimer's disease (AD) through an E.coli model system. We will generate a mutational library of the Amyloid Beta 1-42 (Aβ1-42) gene, to be recombinantly expressed in E.coli as part of a drop-out screen. Given that Aβ1-42 spontaneously aggregates into toxic plaques, we expect the dropout cultures to become enriched over time for Aβ1-42 gene variants correlated with a reduced capacity for aggregation. We will use the recovered information to develop a model identifying key regions of the Aβ1-42 sequence crucial to plaque formation. This can potentially inform future research by revealing plaque-inhibiting Aβ mutations.
WET LAB 2018
Prokaryotic organisms are powerful laboratory tools and form the backbone of synthetic biology. They can be incredibly useful in producing gene products, but also allow us to study the complexities of gene product formation. Producing functional proteins in a cell is dependent on the synthesized protein folding into a unique, thermodynamically stable, structure. However, there are times when proteins mis-fold or unfold and end up forming structures between separate proteins, leading to macromolecular aggregates. Neurodegenerative diseases like Alzheimer’s and Parkinson’s involve the buildup of insoluble protein deposits known as amyloids.
iGEM McMaster’s project this year will be to express human amyloid beta protein in E. coli as a method for studying the molecular basis of aggregation. Prior to insertion, we will randomly mutagenize amyloid beta using error-prone PCR to generate a library. A dropout screen will be used to identify the mutations that are essential for cell viability, with the basis that aggregation due to wildtype amyloid beta is detrimental to the bacterial cells. The screen involves inserting the mutant library into E. coli where it can be expressed. After a period of growth, the sequence distribution of the amyloid beta gene mutant library will be characterized using Next-Gen Sequencing (NGS).
Human Practices 2018
Recent years have seen a rise in concern around neurodegenerative diseases. In 2011, Alzheimer’s Disease (AD) was the seventh leading cause of death in Canada in (1), and the World Health Organization reports that, since 2000, deaths due to dementia have more than doubled throughout the world (2). Our project aims to collect perspectives about AD from community members and researchers. By presenting our work in an accessible video format, we hope to instill a healthy public awareness of AD and initiate wider, better informed discussions surrounding this topic.
Our video will also explore the most celebrated theories of AD prognosis. The amyloid-beta protein, which is being studied in the Wet Lab project, aggregates in AD, yet theories as to whether amyloid-beta is a causative or correlative factor in AD are still largely contested. The Human Practices team will investigate the complexities of AD’s ‘research journey’, with a particular focus on the amyloid-beta hypothesis. Our goal is to understand current theories of AD and current work being done to improve them, while also connecting with and engaging members of the community to better understand their relationship to and perceptions of Alzheimer’s research.
DRY LAB 2018
This year, one of Dry Lab's core projects involves analyzing the data provided by the Wet Lab team. Ideally, this will provide next-generation sequencing data at several time points over the course of their experiments. As the experiments proceed, the sequenced reads should belong to bacteria which better survive the aggregation of plaques. Dry Lab plans to process the reads to identify the most common mutations, how these mutations correlate with each other, and how they become more frequently expressed over time. This should allow the team to identify which mutations affect aggregation, thereby indicating the specific genetic loci of pathogenesis. With enough data, the Dry Lab team can estimate the likelihood of expressing a given mutation based on the presence or absence of other mutations, and we can predict which sequences will confer the greatest survivability.
A main goal for iGEM McMaster is to create lasting initiatives towards public engagement and education around the core ideas and discoveries of our project. In collaboration with Wet Lab and Human Practices teams, the Dry Lab team aims to develop interactive media to share our journey with others, to raise awareness on the issues we aim to tackle, and to inspire and invite others to further our work. As a team, Dry Lab members have discussed the core processes that bely their lab projects, and have conceptualized a series of games that embody their function and intent. In developing analogous narratives around technical scientific processes, the team hopes to break through the seemingly esoteric nature of research into accessible ideas that could engage the public. Throughout development, Dry Lab will engage in dialogue with our target audience to ensure that our initiative continuously aligns with their needs.
Beyond our team's research, we look to apply our depth of mentorship and relationships with the city to facilitate a broader educational outreach strategy, that skillfully leverages our role as students to serve as peer educators in the broader McMaster and Hamilton community. The access to high-quality expertise across the sciences, humanities, and business have supported the growth of a strong culture of mentorship in McMaster’s iGEM team. Consistent with the aims of McMaster University as an institution for higher learning that appreciates both peer-based, inquiry-driven, and student-centred learning, our team has modelled many such approaches towards the cultivation of the next generation of STEM leaders.
Over the past year we have been a part of the annual day-long enrichment program known as Discovery Day, hosted by Hamilton Health Sciences and the Canadian Medical Hall of Fame, attended the GFSS Health Science Symposium, and presented workshops at the Research in Science Exhibition (RISE) conference hosted by the Foundation for Student Science and Technology. We continuously look to invest in young students and to be a part of cultivating the new generation of scientists through mentorship and education.