Core Facilities > Gene Editing Facility

Background
We are establishing a core facility in partnership with Horizon Discovery that enables the manufacture of cell lines where the endogenous gene of interest is mutated. This enables point mutations or deletions to be introduced to replicate known disease causing mutations. These stable cell lines will be isogenic to the parental wild types and will be useful for both basic and translational research applications.

Logistics
We successfully got GMM permission from the local GMAG safety committee and a final submission to the HSE is pending. A room in the Cancer Research Centre has been allocated and funding has been supplied to enable this to be properly equipped. We anticipate that the safety permissions and infrastructure will be in place by January 2012.

Science
We have been working through the preliminary stages of isogenic cell line development. This involves sequencing genomic DNA to ensure that the target sequences are correct for the cell line of choice rather than rely on the reference sequences. This serves two purposes - to ensure optimal homologous recombination and to indicate suitable primers for later QC of your colonies. In collaboration with Horizon Discovery we have designed plasmids for an initial test runs. These will be used to make adeno-associated viruses and then applied to making isogenic cells. It is anticipated that expertise in all of the steps to make isogenic cells will be in place by the end of February 2012.

Current estimates of costs per cell line are £2,000 for the first round of gene editing, targetting the second allele will cost a further £1,000.

Summary
Please contact Ian Prior to discuss your requirements for making isogenic cells. The lab operating model will be that the Gene editing RA will show a member of your group how to perform each of the steps and for the first round at least these steps will be performed in the cell lab in the Cancer Research Centre. Initial discussions can already take place regarding selection of primers for sequencing your genomic DNA and plasmid design.