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A visual representation of DNA editing.

Genome editing

Building experimental models using cutting-edge genetic manipulation.

The Genome Editing Unit (GEU) applies the latest molecular technologies to help generate your research models, from plasmid vectors and cultured cells to whole organisms.

With years of experience applying CRISPR-Cas9, and access to the ever growing molecular toolbox, we work with you to build better, bespoke research models to truly reflect your biological system of interest.

Every project we undertake is different. We provide guidance, and tailor our expertise to develop tools according to your needs.

 

On this page:

 

Liquid being pipetted into an aliquot.
 

How we work

We produce bespoke experimental models, working in collaboration with our research colleagues. We provide training and support to our users.

Services

Our staff are experts in the application of genetic manipulation techniques in a variety of model systems. We advise you on the best way to make your models, and then put it into practice.

The facility is managed full time by an experienced Senior Experimental Officer, and service arms are led by Experimental Officers and Technicians.

Training

We want to ensure user groups take a genetically modified model product from us, and understand how it was made. We offer support and training to our user groups on all aspects of the process, from in silico design through to detecting the desired genetic manipulation

Training is provided in multiple formats, including workshops, seminars, and one-to-one sessions.

 

Applications

The facility has four arms of activity, covering a range of services and support.

Transgenic mice

We offer a complete service from concept to germline confirmed transgenic mouse line.

This includes project design, preparation and purification of reagents, the generation of the transgenic mouse through embryo microinjection, and confirmation of germline transmission.

Typical CRISPR gene edited mouse models include:

  • gene knockouts;
  • genomic deletions;
  • point mutations;
  • conditional alleles;
  • reporter tagged genes;
  • Cre drivers.

We also have experience with a range of alternative transgenesis techniques, including integration of recombinant plasmids and/or bacterial artificial chromosomes (BACs).

We have built viral vectors for use in in vivo, including AAV-Cre and AAV-CRISPR, allowing you to perform genetic manipulations without the need to create a new transgenic line.

Cell line engineering

Exploiting our expertise with CRISPR-Cas9, we have recently added cell line engineering to our service portfolio.

We offer a modular service, where the research group can pay for different aspects of the sequential process according to their budget.

We offer a range of services, including:

  • optimising CRISPR knockout for your cell line of choice;
  • designing and screening gRNA for your target gene knockout;
  • designing CRISPR to create single nucleotide changes;
  • targeted knock-in of larger sequences such as transcriptional reporters, fluorescent fusion genes, or whole transgenes;
  • generating modified cells as either a pooled or clonal population.

We also have expertise in the construction and application of classic overexpression systems, such as plasmid or lentivirus.

We want to ensure the next generation of scientists gain working knowledge of the cutting-edge techniques we apply. We are keen to train staff and students as part of this service.

Plasmid cloning

Our team has years of experience building custom plasmids according to researcher needs, using a variety of molecular biology techniques.

With a library of plasmids and gene tags to choose from, we can build custom genetically encoded constructs for a range of downstream applications, including CRISPR donors and gene expression plasmids, and lentiviral vectors.

We can add functional tags to your genes, including:

  • fluorescent proteins;
  • luminescent reporters;
  • proximity labelling enzymes;
  • pull down epitopes;
  • multifunctional tags (such as HaloTag);
  • inducible protein depletion degron tags.

We also design and clone CRISPR vectors to create genetically modified Drosophila, in collaboration with our colleagues in the Fly Facility.

Research and development

Building on the revolution in molecular technologies, and as the expert hub of gene editing and molecular biology, we have collaborated with local research groups to apply the latest molecular technologies in novel contexts.

We have integrated research and development capacity, and can discuss your project requirements to see if we can assist by building bespoke technologies for specific research goals. Where appropriate, we can collaborate on grant applications.

We have supported a diverse range of technology application and development, including:

  • adapting CRISPR for other model organisms and systems (zebrafish, Drosophila, stem cells);
  • applying gene perturbation technologies such as CRISPRinteference, CRISPRactivation and CRISPR-Cas13;
  • building expression systems with temporal control, such as doxycycline inducible promoters, and controlled protein depletion using Degrons;
  • integrated CRISPR diagnostics for programmable sequence detection.

We have also assisted local groups with the design and development of putative gene and cell therapy approaches for genetic disease.

Gloved hand placing petri dish under a microscope.
 

Publications and outputs

The Genome Editing Unit has been instrumental in research leading to several high impact publications.

Nuclear receptor REVERBĪ± is a state-dependent regulator of liver energy metabolism.

Proceedings of the National Academy of Sciences (PNAS), 2020.

To our knowledge this is the first example of a CRISPR knock-in mouse expressing a HaloTag-gene fusion (Halotag-Reverbα).

This transforms the capacity to perform live single cell imaging combined with dynamic Chip-seq analysis of rhythmically expressed proteins, and allowed us to build an accurate map of the Reverbα cistrome revealing a new role for hepatic Reverbα.

DOI: 10.1073/pnas.2005330117

Analysis of chromatin organization and gene expression in T cells identifies functional genes for rheumatoid arthritis

Nature Communication, 2020.

Genome-wide association studies (GWAS) has identified large numbers of credible disease (psoriasis and arthritis) associated SNPs that reside in non-coding genomic regions.

We assisted with a strategy to functionally probe these sequences, predicted to be enhancers and regulatory regions, using the derivative CRISPRactivation technology.

With this approach, the group has been able to identify genes that are under the transcriptional regulatory control of these putative enhancers.

DOI: 10.1038/s41467-020-18180-7

CRISPR-mediated knock-in in the mouse embryo using long single stranded DNA donors synthesised by biotinylated PCR.

Methods, 2021.

This is a core facility led paper.

There is a pressing need to improve the DNA donors used in CRISPR knock-in in mouse embryo. Increasing the efficiency of gene knock-in would have significant 3Rs impact.

Long ssDNA donors have emerged as optimal donors with increased rates of knock-in via homology directed repair (HDR).

However, present synthesis methods do not produce adequate yields and often contain sequence errors. The method we developed in-house resolves these issues, generating high yield, sequence perfect lssDNA.

As a core facility, we have shared this technique with several local research groups to use in difficult-to-transfect cultured cells and zebrafish embryos.

Reproducibility of CRISPR-Cas9 methods for generation of conditional mouse alleles: a multi-center evaluation.

Genome Biology, 2019. 

Conditional alleles are a popular transgenic mouse model. However, published methods to generate such transgenic mice were shown to be low efficiency when applied at scale.

As a lead centre within this consortium of transgenic core facilities worldwide we contributed CRISPR data from several projects, helping highlight flaws with the current methods for generating conditional alleles in mice, as well as provide data for alternative, more efficient strategies.

Global transcriptomic analysis of the arcuate nucleus following chronic glucocorticoid treatment.

Molecular Metabolism, 2019.

Generating homozygous knockout transgenic mouse strains can be costly and time consuming.

Here, we applied a novel CRISPR gene KO approach to deliver CRISPR reagents directly to the mouse brain via Adeno Associated Virus (AAV).

This allowed us to rapidly and cost effectively validate candidate genes in vivo from ex vivo RNA-seq analysis.

 

External access

Although we primarily support research within The University of Manchester, we are also able to support work from other academic institutions or industry.

Academic institutions

We have generated transgenic mouse models, cell lines and custom vectors for collaborators in academia.

Our capacity for performing external work is dependent on our internal workload and on the complexity of the work to be performed

Industry

We have worked with a number of companies in a variety of capacities.

All work is fully documented and contractual and non-disclosure compliance is assured.

Please note that due to patents and licencing we do not have the freedom to operate with some technologies for industry clients.

If you are interested in accessing the facility please contact our Business Development Manager for further details:

Dr Joanne Flannelly
Email: joanne.flannelly@manchester.ac.uk

96 well plate being prepared for processing.
 

Contact us

Please get in touch if you require any further information, or would like to inquire about using our facility.

Antony Adamson, Senior Experimental Officer

Email: genome.editing@manchester.ac.uk

Genome Editing Unit
Michael Smith Building
The University of Manchester
Oxford Road
Manchester
M13 9PT

Maps and travel

We are based in the Michael Smith Building (Building 71 on the University campus map).

See: University maps and travel section

Technology platforms

We have a pioneering environment and facilities for research, innovation and technology development.

Technology platforms main page

Cross facility support