Transgenic Project

In Transgenic and Genome Editing Core, transgenic mice are generated by pronuclear microinjection of foreign DNA fragments into one-cell-stage mouse embryos. On average, it takes about three to four months to generate a transgenic mouse strain.

The Core can assist investigators with the design of transgenic DNA constructs. Once a transgenic construct has been assembled, the investigator files a request for microinjection services.

Core personnel then excise the transgene from the plasmid backbone and prepare it for microinjection. They also set up superovulation and mating to obtain one-cell-stage embryos for microinjection and prepare pseudopregnant surrogate mothers for embryo-transfer procedures.

The DNA construct is typically injected into fertilized. Microinjected embryos are transferred into surrogate mothers, which develop these embryos to term.

Once pups are born from these eggs, tail biopsies are collected for preparation of genomic DNA. This DNA is screened by PCR analysis for the presence of transgenic DNA. Transgenic animals are then bred to establish a cohort of mice, which can be used to study the phenotypic consequences of transgene expression.

Lin Gan, PhD
Director, Genome Editing Core

Services

 Transgenic DNA microinjection-only service

  • This service includes one microinjection session using C57BL/6J embryos.
  • DNA template will be provided by the users.
  • For each microinjection session, we will inject ≥150 C57BL/6J embryos, perform embryo transfers to pseudopregnant females.
  • Tail tip biopsies will be provided to the investigator's laboratory for genotyping. Mouse pups will be transferred to the investigator at weaning.

 dsDNA synthesis and purification for microinjection

  • DNA template will be synthesized and cloned into plasmid.
  • Mid- or large-preparation of plasmid DNA will be performed and dsDNA fragment will be isolated and purified for microinjection.