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Transgenic/Gene Targeting

The Department of Pathobiology has established a core facility for the production of transgenic and gene targeted mice. The core is composed of three principal elements:

  • Molecular analysis and development of gene constructs for pronuclear injection and gene targeting.
  • Production of transgenic mice by pronuclear injection (overexpression) and blastocyst injection (gene targeting).
  • Detection, analysis and breeding of transgenic mice.

This core facility is supervised by Dr. Ann Kier (1-979-845-3365).

Further core facilities are available in the department for the morphometric analysis of transgenic animals and their tissues. The animals are maintained in a specific pathogen free (SPF) barrier at the Laboratory Animal Resources and Research Facility at Texas A&M University.

Dr. Kier will advise and assist in the development of protocols utilizing the following methodologies:

  • Embryonic stem (ES) cell library screening - mouse
  • ES cell culture
  • Electroporation of ES cells
  • DNA screening by Southern analysis and PCR
  • Superovulation and embryo collection and transfer
  • Pronuclear microinjection
  • Blastocyst microinjection
  • Analysis of sperm by PCR for the evaluation of transgenic males
  • Embryo derivation for the elimination of specific mouse pathogens

Core Equipment

Inverted microscope: A Nikon Diaphot 300 equipped for brightfield, phase contrast, Nomarski differential interference contrast and epi-fluorescence observation. The microscope has both 35 mm still photography and video capabilities.

Micromanipulators: A pair of Narshige MO-188 and MN-188 hydraulic micromanipulators are mounted on the Nikon Diaphot for the holding and injection of embryos.

Inverted microscope table: A gimbal gas piston isolator for vertical and horizontal isolation of vibration.

Pipet Puller: A Stoelting APP-1 microprocessor controlled horizontal pipet puller. This pipet puller is used for the production of injection pipets which can be accurately and repeatedly reproduced.

Microscope: A Nikon TMS inverted microscope equipped for brightfield and phase contrast observation.

Biologic Safety Cabinet: A Nuaire model NU425-400 HEPA filter vertical laminar flow hood is used for tissue culture procedures.

CO2 incubator: A Nuaire model NU-2500 CO2 incubator which is equipped with infrared CO2 monitoring, HEPA filtered air and humidity monitoring, is used for tissue and embryo culture procedures.