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Seminar: Dr Marcus Keatinge, from the University of Edinburgh

Dates:30 May 2025
Times:13:00 - 14:00
What is it:Seminar
Organiser:Faculty of Biology, Medicine and Health
Who is it for:University staff, Current University students
Speaker:Dr Marcus Keatinge
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  • By Faculty of Biology, Medicine and Health

Dr Marcus Keatinge, from the University of Edinburgh, will be giving a seminar on Friday 30 May at 13:00 in the Michael Smith building room B.4208.

Title: “CRISPR-Cas9 allows for highly sophisticated genetic engineering in zebrafish: Knock Outs, Knock Ins and cell type specific humanisation in vivo”

Abstract: Crispr/Cas9 is highly efficient in zebrafish, enabling the acute generation of gene knockouts, which we use for in vivo genetic screens in the zebrafish system. More recently at Edinburgh, knock-in strategies have become more streamlined and reliable for zebrafish. We now routinely insert fluorescent proteins into genomic loci in a targeted manner, facilitating transcriptional reporting and direct protein fusions to track cellular localization. Using the same knock-in strategy, we have also introduced non fluorescent proteins such as NTR2.0 (for targeted cell ablation) and Cre recombinase (for conditional gene manipulation).

Furthermore, we have had considerable experience establishing different methods for cell type-specific knockouts in the zebrafish model, using both Cas9 transgenic approaches as well as Cre-Lox based approaches. Specifically, to remove gene function in the different cells of the CNS in a targeted manner.

We have also adapted a conditional knockout strategy, enabling us to humanize zebrafish genes in a cell type-specific or temporal manner in vivo. This novel approach allows us to convert endogenous zebrafish genes into their diseased human counterparts with remarkable precision. As a result, toxic human proteins associated with neurodegeneration can be expressed in a conditional and physiologically relevant context. Finally, we are currently attempting to insert secondary markers into genes of interest to simultaneously mutate and label them. These would create loss of function alleles which can be genotyped by visual inspection alone, eliminating the need for fin clipping. Together, these advancements highlight the sophisticated level of genetic engineering now achievable in zebrafish.

Short Bio: Dr Marcus Keatinge is a Senior Postdoctoral Research Fellow at Dementia Research Institute - University of Edinburgh. He has vast experience in genetic engineering using CRISPR in zebrafish for acute gene targeting screens to identify modulators of spinal cord repair. His current research is pioneering novel genetic strategies for cell-type-specific humanization in zebrafish to model motor neuron disease in vivo.

Speaker

Dr Marcus Keatinge

Role: Dr

Organisation: University of Edinburgh

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B.4208
Michael Smith Building
Manchester

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Moe Gavin

moe.gavin@manchester.ac.uk

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