Creating Chimaeric Founders
Before any microinjection service can commence a risk assessment for genetically alter animals must be approved by the Biosafety Committee:
For customers who ship ES cells to the facility for microinjection, pathogen testing and karyotype analysis is required prior to injection. Clear instructions as to the growth and selection history of the cells must be outlined prior to shipping the cells. Prior consultation with the facility is required for these protocols.
Pathogen testing is performed by Charles River. The customer can ship the cells to Charles River or any other accredited pathogen screening facility for testing, (IDEXX BioResearch, www.idexxbioresearch.com/radil/radil-EU/index.html) and send GTF paperwork demonstrating that the cells are pathogen-free. GTF offers a service to grow untested ES cell clones in a quarantine hood and incubator and send the samples for a test before generation of chimeras on a cost-recovery basis.
Karyotype analysis, chromosome counting are performed by the GTF.
We will microinject ES cells into approximately 50 blastocysts. Injected blastocysts will be transferred to the uterine horns of pseudopregnant foster mothers and will reside within an isolator in the SOPF section of BSU. Animals (male chimeras) will be screened for germ line transmission by PCR using surrogate females (2) and high transmitting animals will be transferred to the customers project licence at this stage for further colony establishment and expansion.
Chimeric mice will be shipped to the customer at three weeks of age at the customer’s expense.