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CRISPR-Cas9 genome editing and super-resolved live cell imaging to study the extracellular microenvironment at the single molecule level

Project Description

The three-dimensional space around and between mammalian cells is filled with a complex network of extracellular matrix (ECM) molecules, which instruct fundamental processes such as cell survival, differentiation and migration, in addition to providing structural support for cells. ECM molecules orchestrate cell behaviour directly, by signalling to cells, and indirectly, by determining the availability and distribution of secreted proteins such as cytokines and growth factors, and by directing the positioning of cells within tissues. The combination, stoichiometry and geometry of the distinct ECM molecules that are released by cells into their surrounding microenvironment vary from site to site during development, tissue damage, infection and disease.
The aim of this project is to study the secretion, spatial distribution and assembly of the ECM at the single molecule level in real time and how this changes during infection. CRISPR-Cas9 genome editing, super resolution microscopy and physiological 3D cell culture systems will be combined to generate stable cell lines expressing tagged ECM molecules at endogenous levels. This will allow the generation of stable biological models to accurately study endogenous ECM protein stoichiometries, kinetics, spatial relationships and interactions. There is also the possibility to investigate the release of ECM molecules by non-canonical cell sources, including macrophages, in order to understand its functional implications, which are currently unknown.

Laboratory Environment and Training Opportunities
The project will be carried out within the Gene Regulation & RNA Biology Group (Division of Biomolecular Science & Medicinal Chemistry) which currently consists of 6 principal investigators and around 25 full time researchers with well-equipped laboratory facilities on the main campus. The School of Pharmacy provides a vibrant cross-disciplinary research environment, with excellent support and training facilities for PG students. The student will have access to a wide range of different sources of training, both in advanced scientific techniques and transferable skills. The student will attend division and PG meetings and will be expected to attend seminars within the School and those relevant in the wider University. Subject-specific training will be received through our group’s regular supervision meetings.

Funding Notes

Applications are welcome from motivated students with Bachelors (2.1 or above) or Masters degree(s) in Science subjects. International applicants should meet the University requirements for English. Prospective candidates should visit our University pages for information regarding fees and funding at the University. Sponsored and self-funded students are encouraged to contact the supervisor for further information ().


1) ANNA M. PICCININI, LORENA ZULIANI-ALVAREZ, JENNY M. P. LIM and KIM S. MIDWOOD, 2016. Distinct microenvironmental cues stimulate divergent TLR4-mediated signaling pathways in macrophages Science Signaling. 9(443), ra86
2) TOMLIN H and PICCININI AM, 2018. A complex interplay between the extracellular matrix and the innate immune response to microbial pathogens. Immunology. 155(2), 186-201
3) GOH, F. G., PICCININI, A. M., KRAUSGRUBER, T., UDALOVA, I. A. and MIDWOOD, K. S., 2010. Transcriptional regulation of the endogenous danger signal tenascin-C: a novel autocrine loop in inflammation: J Immunol J Immunol. 184(5), 2655-62

How good is research at University of Nottingham in Allied Health Professions, Dentistry, Nursing and Pharmacy?

FTE Category A staff submitted: 44.10

Research output data provided by the Research Excellence Framework (REF)

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