Molecular regulation of cell behaviour during muscle regeneration and disease.
The Knight group aims to understand how cell behaviour is related to tissue regeneration and stem cell function with a focus on muscle. We use a variety of advanced imaging approaches to observe and manipulate cells in vivo (zebrafish), ex vivo (mouse) or in vitro (human) in order to develop complex models of gene function and cell behaviour. These allow us to make predictions about gene function in the control of muscle, immune and connective cells during tissue repair.
In the video below, Dr. Knight explains how he aims to identify genes and molecules that control how cells behave during regeneration in order to inform the development of new therapies for tissue repair. He uses a powerful combination of live cell imaging and molecular genetics to test how specific genes control immune and stem cells to induce healing.
Publications
Awards
BBSRC project award
MDUK studentship
NC3Rs studentship
Royal Society International Partnership Award
Carl Zeiss collaborative award
Activities
Meetings
At the recent Gordon Myogenesis meeting held in Tuscany June 2019, Robert presented the latest results from the group, demonstrating the link between cell migration and proliferation as muSCs respond to injury in zebrafish. Camilla, a former student in the group, attended the FSHD international conference in Marseille, June 2019. She presented her work in developing a zebrafish model of FSHD and results showing how the FSHD associated gene, DUX4, affects muscle regeneration.
Collaboration
A joint project with Professor Ivo Sbalzarini at the Centre for Systems Biology in Dresden aims to develop powerful computational tools for analysing movies of cells during regeneration. As part of this collaboration, Jsu-Szu Chen, a Masters student from TU Dresden will join the Knight group at King’s to work with the team for 6 months. Funding: TU Dresden - International Strategic Partnership Award King’s College London – International Partnership Award
Collaboration
An ongoing collaboration with Professor Richard Jaspers at VU in Amsterdam, the Netherlands, aims to combine imaging with physiology to determine how muSCs are regulated in vivo. Combining the advanced imaging capacity of the Knight group at King’s with Prof. Jasper’s expertise in physiology the two groups have recently described how muSCs respond to shear force in vivo with implications for their response to tissue injury and repair. Funding: Royal Society International Partnership Award
Collaboration
Working with Professor Benedicte Chazaud at the CNRS in Lyon, France, we have been investigating the function of a novel factor secreted by macrophages that her group discovered. Using transgenic zebrafish and live cell imaging we have shown how over-expression of this factor can alter muSC function leading to changes in proliferation and fusion during regeneration. Funding: Royal Society International Partnership Award
ZEISS Multiphoton Microscope
Multiphoton microscopy is a laser-scanning technique that employs non-linear effects at the objective focus to create optical sectioning. Longer (near-infrared) wavelengths are used which minimise photodamage and significantly increase the penetration depth, allowing 3D volumetric imaging at greater depths (hundreds of microns) into tissue than confocal microscopy. Installed in 2015, this bespoke multiphoton microscope allows multicolour imaging with up to three excitation wavelengths and eight detection channels.
ZEISS 7MP with Ti:sapphire laser and MPX for dual-excitation and extended wavelength imaging
Specifications
- Broad range of imaging wavelengths from 690-1340 nm. - Two laser lines, allowing simultaneous...
Objectives available
- 10 x, 0.3 NA air - 40 x, 1.3 NA oil immersion - 32x, 0.85 NA water immersion with...
Image analysis
- Two high-specification dedicated image analysis workstations are available to microscope users. - ...
.
Multiphoton Microscope Specialist
Dylan Herzog has extensive knowledge in fluorescence microscopy, especially in the use of multiphoton microscopes. He obtained his PhD on 'Rapid Fluorescence Amplified Biofilm Detection' at King's College London. As well as managing training and access to the microscope, Dylan can also provide expertise in multiphoton imaging and advise on image analysis.
Please feel free to contact dylan.b.herzog@kcl.ac.uk for more information about the microscope.
Useful information on two-photon excitation and emission:
ThermoFisher - Fluorescent Probes for Two-Photon Microscopy: General information and table of excitation wavelengths.
https://www.thermofisher.com/uk/en/home/references/molecular-probes-the-handbook/technical-notes-and-product-highlights/fluorescent-probes-for-two-photon-microscopy.html
Two-photon absorption properties of fluorescent proteins: two-photon absorption/excitation properties of a wide variety of fluorescent proteins
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4772972/pdf/nihms-761967.pdf
Two-photon fluorescence absorption and emission spectra of dyes relevant for cell imaging: Absorption/excitation and emission spectra for fluorophores relevant in cell imaging.
Excitation Spectra and Brightness Optimization of Two-Photon Excited Probes: Two-photon molecular brightness spectra for a diverse set of organic and genetically encoded probes.
https://www.sciencedirect.com/science/article/pii/S000634951200063X?via%3Dihub
Publications
Awards
BBSRC project award
MDUK studentship
NC3Rs studentship
Royal Society International Partnership Award
Carl Zeiss collaborative award
Activities
Meetings
At the recent Gordon Myogenesis meeting held in Tuscany June 2019, Robert presented the latest results from the group, demonstrating the link between cell migration and proliferation as muSCs respond to injury in zebrafish. Camilla, a former student in the group, attended the FSHD international conference in Marseille, June 2019. She presented her work in developing a zebrafish model of FSHD and results showing how the FSHD associated gene, DUX4, affects muscle regeneration.
Collaboration
A joint project with Professor Ivo Sbalzarini at the Centre for Systems Biology in Dresden aims to develop powerful computational tools for analysing movies of cells during regeneration. As part of this collaboration, Jsu-Szu Chen, a Masters student from TU Dresden will join the Knight group at King’s to work with the team for 6 months. Funding: TU Dresden - International Strategic Partnership Award King’s College London – International Partnership Award
Collaboration
An ongoing collaboration with Professor Richard Jaspers at VU in Amsterdam, the Netherlands, aims to combine imaging with physiology to determine how muSCs are regulated in vivo. Combining the advanced imaging capacity of the Knight group at King’s with Prof. Jasper’s expertise in physiology the two groups have recently described how muSCs respond to shear force in vivo with implications for their response to tissue injury and repair. Funding: Royal Society International Partnership Award
Collaboration
Working with Professor Benedicte Chazaud at the CNRS in Lyon, France, we have been investigating the function of a novel factor secreted by macrophages that her group discovered. Using transgenic zebrafish and live cell imaging we have shown how over-expression of this factor can alter muSC function leading to changes in proliferation and fusion during regeneration. Funding: Royal Society International Partnership Award
ZEISS Multiphoton Microscope
Multiphoton microscopy is a laser-scanning technique that employs non-linear effects at the objective focus to create optical sectioning. Longer (near-infrared) wavelengths are used which minimise photodamage and significantly increase the penetration depth, allowing 3D volumetric imaging at greater depths (hundreds of microns) into tissue than confocal microscopy. Installed in 2015, this bespoke multiphoton microscope allows multicolour imaging with up to three excitation wavelengths and eight detection channels.
ZEISS 7MP with Ti:sapphire laser and MPX for dual-excitation and extended wavelength imaging
Specifications
- Broad range of imaging wavelengths from 690-1340 nm. - Two laser lines, allowing simultaneous...
Objectives available
- 10 x, 0.3 NA air - 40 x, 1.3 NA oil immersion - 32x, 0.85 NA water immersion with...
Image analysis
- Two high-specification dedicated image analysis workstations are available to microscope users. - ...
.
Multiphoton Microscope Specialist
Dylan Herzog has extensive knowledge in fluorescence microscopy, especially in the use of multiphoton microscopes. He obtained his PhD on 'Rapid Fluorescence Amplified Biofilm Detection' at King's College London. As well as managing training and access to the microscope, Dylan can also provide expertise in multiphoton imaging and advise on image analysis.
Please feel free to contact dylan.b.herzog@kcl.ac.uk for more information about the microscope.
Useful information on two-photon excitation and emission:
ThermoFisher - Fluorescent Probes for Two-Photon Microscopy: General information and table of excitation wavelengths.
https://www.thermofisher.com/uk/en/home/references/molecular-probes-the-handbook/technical-notes-and-product-highlights/fluorescent-probes-for-two-photon-microscopy.html
Two-photon absorption properties of fluorescent proteins: two-photon absorption/excitation properties of a wide variety of fluorescent proteins
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4772972/pdf/nihms-761967.pdf
Two-photon fluorescence absorption and emission spectra of dyes relevant for cell imaging: Absorption/excitation and emission spectra for fluorophores relevant in cell imaging.
Excitation Spectra and Brightness Optimization of Two-Photon Excited Probes: Two-photon molecular brightness spectra for a diverse set of organic and genetically encoded probes.
https://www.sciencedirect.com/science/article/pii/S000634951200063X?via%3Dihub
Group lead
Robert Knight
Reader in Developmental Genetics
Contact us
Dr Robert Knight
Centre for Craniofacial Development & Regenerative Biology
Floor 27, Guy's Tower WingGuy's Hospital