Table of Services

Transgenic Mouse Facility

Description

The facility occupies two newly renovated rooms, the first of which contains two microinjection work stations, one for DNA microinjection, and the other for injection of blastocysts. The second room is colony space that houses the progenitor strains and the transgenic mice during gestation until reproductive maturity, when they are transferred to the investigator's colony. The animals are housed in microisolator cages, which are changed inside laminar flow hoods. Both rooms are supplied with HEPA- filtered air and designed with air pressure positive to the hallway to maintain a clean environment.

The Transgenic Mouse Facility produces transgenic and gene-targeted mice, and rederives existing mouse lines.

Transgenic mice are produced by pronuclear injection of a gene construct, provided by a Center investigator, into mouse zygotes, which develop to term after surgical transfer into recipient female mice. After weaning, tissue samples from the offspring are given to the investigator for genotyping. Mice in which the transgene has integrated into the genome are identified by the investigator's laboratory using PCR or Southern blot analysis. Positive "founder" mice are transferred to the investigator for breeding and experimental analysis of transgenic lines.

Gene-targeted mice are produced by microinjecting embryonic stem (ES) cells, which have been genetically altered through the efforts of Center investigators and/or the Cell Culture Facility, into blastocyst stage mouse embryos. After surgical transfer and development to term, mice which carry the engineered ES cells are identified by coat color and transferred to the investigator for breeding and analysis.

Mouse lines are rederived using embryo transfer techniques to generate pathogen- free mice from progenitors which are infected with murine pathogens.

The Transgenic Facility provides assistance to the investigator in such areas as construct design, analysis of transgenic mice, and transgenic breeding schemes.