The Transgenic/Knock-out Mouse Facility offers the services shown below. If you are a Massey member, the cost of these services is partially subsidized as a benefit of membership.
Transgenic / Knock-out services
- Transgenic mouse production: The core generates at least three founder mice for each transgene.
- Knock-out/knock-in mouse production: The core can perform all or a subset of the following procedures:
- ES cell electroporation,
- ES cell culture under appropriate selection conditions,
- expansion of drug-resistant clones,
- PCR/Southern blot screening of clones for the correct recombination event,
- re-electroporation with a cre expression vector to delete floxed selection markers and subsequent re-screening of clones;
- injection of targeted ES cell clones into blastocysts for chimeric mouse production. For projects in which the core has performed the ES cell targeting, at least two independent ES cell clones are generally injected into blastocysts, and a minimum of five chimeras are generated.
- Mouse line rederivation: Lines of pathogen-infected mice are rederived by embryo transfer to generate specific pathogen-free mice.
- Sperm cryopreservation: Lines of mice are cryopreserved by collection, freezing and long-term liquid nitrogen storage of sperm. When needed, re-establishment of the line can be accomplished by thawing sperm and performing iIVIS Spectrumn vitro fertilization of eggs isolated from WT females.
- Tail DNA genotyping: The core isolates genomic DNA from tail biopsies (or other tissue) provided by the investigator and performs PCR genotyping. Consulting services: The core provides consultation on many aspects of transgenic/knock-out mouse technology, including experimental design, vector design, budgeting for mouse colony maintenance and preparation of applications to the IACUC for the use of laboratory animals in research. In addition, the core staff provides instruction on overall colony management, including breeding strategies, weaning, ear punching and tail DNA isolation.
- The core laboratory also maintains an IVIS Spectrum instrument (shown on the right) for live-animal imaging studies.
Note that rates current as of 20 May, 2011, but are subject to change. Massey members pay 75 percent of these rates per the member subsidy program.
- Transgenic mouse production: Investigators are charged $3,000 per construct for the production of transgenic mice. An additional $500 is charged for production of transgenic mice in a C57Bl/6 or other inbred background.
- Knock-out/knock-in mouse production: The cost for the three stages of knock-out/knock-in mouse production is:
- Stage 1 - ES cell electroporation: $3,000 per targeting vector construct; $1,500 for cre expression vector re-electroporation.
- Stage 2 - screening of ES cell clones: $2,000 for three 96-well plates; $1,000 for screening after cre expression vector re-electroporation.
- Stage 3 - blastocyst injection: $3,000 for two clones.
- Mouse line rederivation: The charge for rederivation of a line of mice by embryo transfer is $1,000.
- Sperm cryopreservation: The charge for cryopreservation of sperm from two males of a single line and long-term storage is $750
- Tail DNA genotyping: The core charges $5 per sample for genomic DNA purification and an additional $5 per sample for PCR genotyping.
- Consulting/support services: There is no charge for initial consultations.
As of January 1, 2012, VCU Massey subsidizes 25 percent of the cost of members’ core laboratory usage for the following Massey-supported, fee-for-service shared resources:
- Biological Macromolecule Shared Resource Core
- Flow Cytometry Shared Resource Core
- Structural Biology Shared Resource Core
- Tissue and Data Acquisition and Analysis Core
- Transgenic/Knock-out Mouse Facility
Note that this subsidy is available only for Massey members. To receive this member benefit, please initiate core lab services using a VCU index code associated with your name. When you are billed for core laboratory usage, the CORES system will automatically split the invoice between your index code and a Massey account established for this purpose. You will be charged 75 percent of the published rate and VCU Massey will provide a 25 percent co-pay to cover the balance.
Please direct any questions or concerns about the member subsidy benefit to Paul Fawcett, Ph.D. at firstname.lastname@example.org or 804-833-1959.