Introduction

The Genome Engineering Facility at the WIMM is a central core service set up to establish genetically modified model systems and is available to all researchers in the units. We use conventional cloning and recombineering approaches to generate DNA-based targeting constructs such as large BAC-transgenes, compound transgenic constructs and complex targeting constructs, such as conditional knock-out (cKO), conditional knock-in (cKI) as well as customer-defined transgenic cassettes. For targeting your cell line or species of choice, we mainly use the CRISPR/Cas9 technology platform. Insertion of larger cassettes in mouse models is performed using Cas9-assisted homologous recombination in mouse ES cells. The aim and expertise of our Genome Engineering service is to advise on the experimental strategy to choose, and upon agreement, to design and assemble any kind of genetic targeting construct and ready-to-use validated CRISPR reagents.

 

fig1
Figure 1: Genome Engineering using the type II CRISPR/Cas9 system of S. pyogenes. Single short guide RNAs (sgRNAs) can target the prokaryotic Cas9 DNA-endonuclease to virtually any given spot in the genome of your target organism. After the cut, double strand breaks can be repaired by the target cell by distinct repair pathways. In presence of a repair template, Homology Directed Repair (HDR) can be applied, in absence of a donor the cell is bound to use the efficient Non-Homologous End Joining (NHEJ) pathway. Figure modified from (Charpentier et al., Nature, 2013).

Services we provide are detailed below, and you can find out more information by following the indicated hyperlinks. We use a wide range of molecular biology techniques and RedET recombineering for the assembly of DNA-based targeting constructs. We offer a CRISPR/Cas9 pipeline for the modification of mouse and human cell lines and design and generate reagents used for the generation of animal models. Especially for mouse mutagenesis, together with the WIMM transgenic (TG) services we advise on pronuclear injection strategies for mouse oocytes and, as a recent addition to our portfolio, we offer a Cas9-assisted mouse embryonic stem cell targeting pipeline for the generation of genetically modified mouse models. The ESC-targeting is done in collaboration with the WTCHG Core Transgenics Facility, blastocyst injections will be performed by the WIMM TG facility.

In our internal download section, we provide a variety of CRISPR/Cas9 relevant protocols, general information on the establishment of clonal cell lines, general screening information and our WIMM-internal price list. Whilst all consultation and most design processes are free, we do operate on a cost recovery basis and have a minimal service charge to recover consumables and part of the staff expenses. However, we try to keep our prices on the low side to make our services affordable.

You can find a selected list of publications in which model systems generated by the WIMM Genome Engineering Services have been successfully used, and we are looking forward to make this list much longer.

We also offer our services to researchers in other departments of Oxford University. If you have a project in mind and would like to discuss the details, require advice on genome engineering issues and molecular cloning technology, or need a detailed quote for your specific project, please contact us to find out what we can do for you!

Meet the team:     

Philip Hublitz (philip.hublitz@ndcls.ox.ac.uk), Head of Genome Engineering.
Zoe Holloway (zoe.holloway@ndcls.ox.ac.uk), Research Assistant.
Phone: +44 (0) 1865 2-22339.
Fax: +44 (0) 1865 2-22500.
Location: WIMM, 1st floor, room 343.