Microtubule organisation during gut organoid morphogenesis
Lead Research Organisation:
University of East Anglia
Department Name: Biological Sciences
Abstract
Polarisation and differentiation of columnar epithelial cells is critical for normal gut function and defects in polarisation lead to loss of gut barrier function, epithelial cell invasion and cancer. Importantly, polarised elongation of epithelial cells is dependent on microtubule reorganisation into apico-basal arrays. However, the molecules and mechanisms responsible for this remain to be determined. The aims of this PhD studentship are to use 3D in vitro gut organoids that mimic in vivo gut development to study the role of novel microtubule assocciated proteins in the generation, stability and function of apico-basal microtubule arrays during epithelial differentiation. GFP, RNAi and CRISPR/Cas9 technology in conjunction with live time-lapse, confocal, multiphoton, super-resolution and electron microscopy will be used to pursue these aims. This study presents an exciting opportunity to elucidate the molecular mechanisms responsible for microtubule reorganisation that underpins epithelial differentiation and tissue formation and function.
Organisations
People |
ORCID iD |
| Mette Mogensen (Primary Supervisor) | |
| Benjamin Rix (Student) |
Studentship Projects
| Project Reference | Relationship | Related To | Start | End | Student Name |
|---|---|---|---|---|---|
| BB/M011216/1 | 01/10/2015 | 31/03/2024 | |||
| 1800048 | Studentship | BB/M011216/1 | 01/10/2016 | 31/03/2021 | Benjamin Rix |
| Description | I have furthered the understanding of the role of the centrosome in the process of epithelial differentiation. The centrosome appears to be essential for the formation of 3D cysts but not essential for the formation of polarised 2D layers. I have also studied the microtubule binding protein EB2, which is highly expressed in some undifferentiated cells and also in some cancers. This work includes analysis of the effect of EB2 expression on cell morphology and in particular the behaviour of the centrosome. Ultimately this work answers questions on the role of EB2 in differentiation and polarisation of epithelial tissues. |
| Exploitation Route | EB2 is a very understudied protein but clearly is very important to many cellular processes as it influences microtubule behaviour which has many downstream effects. Further study of EB2 may lead to the development of therapeutic targets for many diseases. |
| Sectors | Pharmaceuticals and Medical Biotechnology |
| Title | CRISPR |
| Description | Generation of an EB2 knock out cell line using the CRISPR technique in combination with the insertion of a selection marker. |
| Type Of Material | Cell line |
| Year Produced | 2020 |
| Provided To Others? | No |
| Impact | Generation of an EB2 knock out cell line will be very important for further research in this lab, allowing many more observations to be made about the role of EB2. |
| Description | Associate Tutor |
| Form Of Engagement Activity | Participation in an activity, workshop or similar |
| Part Of Official Scheme? | No |
| Geographic Reach | International |
| Primary Audience | Undergraduate students |
| Results and Impact | Tutoring of undergraduate and postgraduate students who were taught the indirect immunolabelling technique including imaging, using a fluorescent microscope. |
| Year(s) Of Engagement Activity | 2017,2018,2019,2020 |
| Description | Hosting a visit for CEOs and fundraisers of local charities |
| Form Of Engagement Activity | Participation in an activity, workshop or similar |
| Part Of Official Scheme? | No |
| Geographic Reach | Local |
| Primary Audience | Supporters |
| Results and Impact | Members of local cancer charities that provided funding for new equipment used as part of this research were introduced to the research and techniques carried out by our lab. They were then given a tour of the facilities to experience and appreciated the level of research taking place. |
| Year(s) Of Engagement Activity | 2019 |
| Description | Hosting high school students thinking about going to college. This included theory of research and lab tours. |
| Form Of Engagement Activity | Participation in an activity, workshop or similar |
| Part Of Official Scheme? | No |
| Geographic Reach | Local |
| Primary Audience | Schools |
| Results and Impact | A tour of the lab and imaging facilities was done including the imaging of cells on a confocal microscope. |
| Year(s) Of Engagement Activity | 2017 |
| Description | Student Supervision |
| Form Of Engagement Activity | Participation in an activity, workshop or similar |
| Part Of Official Scheme? | No |
| Geographic Reach | International |
| Primary Audience | Undergraduate students |
| Results and Impact | Undergraduate and postgraduate students carrying out their research projects were supervised. This included all aspects of research, from lab techniques to analysis and presentation of results. |
| Year(s) Of Engagement Activity | 2017,2018,2019,2020 |