Programming the Notch response: optogenetic strategies to manipulate pathway activity
Lead Research Organisation:
University of Cambridge
Department Name: Physiology Development and Neuroscience
Abstract
Theme: Understanding the rules of life
The Notch pathway is one of a small handful of cell signalling pathways that coordinate development, regulating the types and numbers of cells formed in many developmental contexts. Its functions include maintenance of stem/progenitor cells, regulation of cell fates, organizing patterns of growth, and many others. In addition, aberrant Notch activity is implicated in diseases including cancers. A major focus of the work in the lab is on understanding how the Notch pathway operates and what enables its different functional outcomes.
Recently we have developed methods that allow us to follow specific target genes in living cells making it possible to investigate nuclear events including how the transcription factor dynamics and the chromatin context influences the selection of gene targets. A remaining challenge is how to activate Notch with high precision. To achieve this we will adopt optogenetic methods to release an active form of the Notch receptor, by fusing it to light sensitive domains. Ultimately, these optogenetic tools, combined with methods to track the behaviour of other components in the pathway, will be used to generate accurate models of how the active Notch operates inside a cell to target the correct genes and how this can be disrupted by disease causing modifications.
The Notch pathway is one of a small handful of cell signalling pathways that coordinate development, regulating the types and numbers of cells formed in many developmental contexts. Its functions include maintenance of stem/progenitor cells, regulation of cell fates, organizing patterns of growth, and many others. In addition, aberrant Notch activity is implicated in diseases including cancers. A major focus of the work in the lab is on understanding how the Notch pathway operates and what enables its different functional outcomes.
Recently we have developed methods that allow us to follow specific target genes in living cells making it possible to investigate nuclear events including how the transcription factor dynamics and the chromatin context influences the selection of gene targets. A remaining challenge is how to activate Notch with high precision. To achieve this we will adopt optogenetic methods to release an active form of the Notch receptor, by fusing it to light sensitive domains. Ultimately, these optogenetic tools, combined with methods to track the behaviour of other components in the pathway, will be used to generate accurate models of how the active Notch operates inside a cell to target the correct genes and how this can be disrupted by disease causing modifications.
People |
ORCID iD |
| Sarah Bray (Primary Supervisor) | |
| Jonathan Townson (Student) |
| Title | Podcast with the Naked Scientists |
| Description | I wrote and spoke about using flies as a model organism for "The Naked Scientists" podcast at a "Communicating your Science" workshop with the Genetics Society. |
| Type Of Art | Creative Writing |
| Year Produced | 2018 |
| Impact | It is available to this day! And trained me in Science communication. |
| URL | https://www.thenakedscientists.com/podcasts/short/genetics-society-2018-frenetic-genetic |
| Title | BLITz optogenetics in Drosophila |
| Description | An aim for the grant is to create an optogenetic construct for Notch signalling activation. One potential method is to use BLITz which has been previously used in cells and mice. I have successfully used this in Drosophila. It was found to require some adaptations for use in Notch signalling in order to counter mislocalisation of transcription factors and this has been overcome. |
| Type Of Material | Model of mechanisms or symptoms - non-mammalian in vivo |
| Year Produced | 2019 |
| Provided To Others? | No |
| Impact | It raised further questions for us on the binding of Notch intracellular domain to it's transcription co-factors. This led to us talking over new ideas with Rhett Kovall and designing new experiments. |
| Description | Rhett Kovall |
| Organisation | University of Cincinnati |
| Country | United States |
| Sector | Academic/University |
| PI Contribution | We have a Notch optogenetic construct we were looking to improve by obscuring a binding site for a transcription co-factor |
| Collaborator Contribution | Rhett had the knowledge to tell us which residues to obscure and further we worked on designing an experiment to expose the sites in separate constructs. |
| Impact | New constructs have been made and are still being tested |
| Start Year | 2019 |
| Description | BAME school outreach progamme |
| Form Of Engagement Activity | A talk or presentation |
| Part Of Official Scheme? | No |
| Geographic Reach | Regional |
| Primary Audience | Schools |
| Results and Impact | Clare college Cambridge invited students from target areas in London (e.g. Tower Hamlets) who are part of the BAME community to come to the college for an open day and to hear more about studying at the University of Cambridge. As part of this I have delivered mock supervisions to the students on plant gene editing and cancer immunotherapy (for medics). |
| Year(s) Of Engagement Activity | 2020 |
| Description | No slides talk to postgraduate community |
| Form Of Engagement Activity | A talk or presentation |
| Part Of Official Scheme? | No |
| Geographic Reach | Local |
| Primary Audience | Postgraduate students |
| Results and Impact | I gave a talk on my research, spending a lot of time on using flies as a model organism. The talk was done without slides to a postgraduate community of mixed backgrounds across the Sciences and Arts. |
| Year(s) Of Engagement Activity | 2019 |
| Description | Norfolk council schools higher aspirations scheme |
| Form Of Engagement Activity | A talk or presentation |
| Part Of Official Scheme? | No |
| Geographic Reach | Regional |
| Primary Audience | Schools |
| Results and Impact | Over one hundred year 12 students in Norfolk are participating in a higher aspiration scheme being run by the council and Gonville and Caius college Cambridge. I provide a one hour session every month for six months introducing them to further study/ideas in the field of Biology that they do not learn in school, the idea being to replicate the supervision type learning environment they would get if they successfully applied to the University of Cambridge and encourage them to do so. The title of my supervision series is "Molecule of life: how researchers are studying and using DNA". |
| Year(s) Of Engagement Activity | 2020,2021 |
| Description | School visit (Warwick) |
| Form Of Engagement Activity | Participation in an activity, workshop or similar |
| Part Of Official Scheme? | No |
| Geographic Reach | Regional |
| Primary Audience | Schools |
| Results and Impact | Year five students from across Warwickshire identified as being high achieving congregated in a single school and were introduced to new topics by Cambridge University researchers, one of which was me. I introduced them to DNA as being the "recipe of life" based on the BBSRC engagement activity |
| Year(s) Of Engagement Activity | 2020 |
| URL | https://bbsrc.ukri.org/engagement/schools/keystage3/discovering-dna/ |