Transgenic mouse production

Vivarium and Transgenics Core

Service overview

Transgenic mice are produced by microinjecting DNA constructs into the pronuclei of 0.5 days post coitum (DPC) one-cell fertilized eggs. The injected eggs are transferred to the oviducts of 0.5 DPC pseudopregnant foster mothers. The DNA construct will integrate randomly into the egg’s genome and the resulting founder mice will contain one or more copies of the transgene in every cell. Small tail tissue samples are isolated from each mouse for DNA genotype analysis. Mice determined to be transgenic for the injected DNA are shipped to the client. For pricing, please contact Audra Guikema.

Please prepare the appropriate DNA construct prior to shipment to VARI.

Once the construct is received, Core personnel will complete the following:

DNA microinjection

Staff will microinject purified, linearized DNA into one-cell fertilized eggs. These eggs are obtained by mating (C57BL/6 X C3H)F1 or (C57BL/6 X SJL)F1 females to (C57BL/6 X C3H) F1 or C57BL/6 X SJL)F1 males, respectively. Eggs obtained from other strains of mice are available upon consultation with the Vivarium and Transgenics Core. Once the eggs are injected, the majority are implanted into pseudopregnant foster mothers that same day. A portion will be incubated overnight to assess the construct’s toxicity. Eggs dividing to the two-cell stage will be implanted.

DNA injection is prioritized by the DNA sample’s receipt date; however, clients submitting multiple samples will have one construct injected per week to prevent a backup of samples from other clients. Clients will be notified by email when their constructs have been microinjected. For efficient scheduling, please provide the Vivarium and Transgenics Core with advance notice of sample submissions.

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