Policies

The Neurotransgenic Laboratory operates on a Fee for Service basis. Project scheduling can be discussed with the Director of the Neurotransgenic Facility, Dr. Guoping Feng, and the Lead Scientist, Dr. Bernd Gloss. Before the start of a project, clients are asked to download the required forms ("Start of Project"; "Authorization"; "Cost of Service") from this website, fill in the neccessary information and return the signed forms to the Business Manager (Li Qiu, on the 4th floor of the Bryan Research Building).

The Institutional Biosafety Committee (IBC) also requests that each genetically altered mouse be registered at IBC before we begin the project. The project will then be assigned an IBC registration number which needs to be recorded on form "Start of Project". Please visit the IBC website at http://www.safety.duke.edu/LabSafety/DNA.asp and download the "Recombinant DNA Registration Form".

The construction of a targeting vector and the homologous recombination of the Bacterial Artificial Chromosome (BAC) can be achieved in a forseeable timeframe. However, the process of transgenesis, due to the expression of an exogenous gene product in the fertilized oocyte, cannot be guaranteed. Our facility will inject one, and if necessary another, independent construct (Plasmid or BAC based) into the pronucleus of the recipient oocytes. If no transgenic offspring can be obtained from both independent injections, the transgene would be considered incompatible with embryogenesis and the project would have to be terminated. A fee for one injection would have to be charged to the initiating investigator.

DNA for microinjection will be prepared by the Neurotransgenic lab. Clients who generate their own transgenic vectors are asked to provide single bacterial colonies harboring the transgenic construct on an agar plate. For non-standard transgenes, PCR primer pairs will be requested by the Neurotransgenic lab for confirmation of BAC clones and for later screening of transgenic offspring.

Transgenic offspring will be genotyped by the Neurotransgenic laboratory and housed until weaning. For breeding of the F1 generation (usually to C57Bl6 mice) the transgenic founder animals will be transfered to the client's mouse facility. Investigators have to provide their approved protocol number, location of their mouse room, and rack number if specified, to Jimmy Gross (gross@neuro.duke.edu).

Currently we do not culture embryounic stem cells for homologous recombination. We do, however, provide the service for injection of genetically modified mouse embryonic stem cells into blastocysts.

Because the quality of the embryonic stem cells is essential for the success of the experiment, we rely on the client to provide high quality ES cells. We will inject one or two independent ES cell clones into blastocysts and house the chimeric offspring until weaning and coat color can be detected. Should no coat color contribution by the ES cells be detectable, we would conclude that the injected ES cells did not contribute to the animal's tissues. Though the reasons for that are not always clear, we will have to charge the investigator for one blastocyst transfer at this point.