Mouse Genetics Facility

Services

The Mouse Genetics Facility provides the following services, which are described below in greater detail:

• The generation of transgenic mice by pronuclear injection of transgene constructs
• The generation of knockout/knockin mice by injecting gene-targeted ES cells into blastocysts
• The establishment of gene-targeted ES cell clones by homologous recombination

In addition, the following services are currently being developed:

• The generation of knockout/knockin mice by tetraploid complementation
• Reprogramming and mouse cloning by nuclear transfer
• Lentivirus-based transgenesis
• The generation of siRNA knockdown mice

For billing purposes, Facility services generally fall into three categories: Essential, Full, and Optional.

• Essential services. These include microinjecting transgene constructs or ES cells into blastocysts to produce transgenic mice.
  
• Full services. These include Essential services plus gene targeting in ES cells, genotyping mice, or other combinations of service.
  
• Optional services. These include the use of any technology currently being developed by the Facility (see above). Costs for Optional services will be assessed on a collaboration basis, as opposed to fee-for-service.

The generation of transgenic mice by pronuclear injection of transgene constructs

Pronuclear injection of transgene constructs into zygotes is a reliable and efficient technique for producing transgenic mice. The Facility will use this method to produce and deliver at least two independent transgenic mouse founders per transgene construct. As their mouse strain, investigators may choose either B6CBAF1 (hybrid) or FVB/N (inbred). If you need a strain other than one of these, please contact the Facility Director.

Applications. Transgenic mice are valuable animals to study the phenotypic effects of transgene expression within the context of a whole organism. They offer insight into the temporal and spatial patterns of mammalian gene regulation and expression during organism development, maturation, and pathology. And, when combined with an inducible approach, transgenic mice afford relatively precise control over where and when transgene expression will occur.

To request this service, click the “How to obtain services” link.

To learn how to prepare your transgene constructs for injection, click the “Helpful information” link.

The generation of knockout/knockin mice by injecting gene-targeted ES cells into blastocysts

The Facility delivers chimeras produced by injecting gene-targeted ES cells into blastocysts. These ES cells can be either isolated in the investigator’s laboratory or established by the Facility for an additional fee. Depending on the genetic origin of the ES cells, the genetic background of the Facility’s blastocyst donors will be either C57BL/6 or BALB/C. If you need a strain other than one of these, please contact the Facility Director.

Technology. The injection of gene-targeted ES cells into blastocysts is a reliable and efficient way to produce chimeras, which can then be backcrossed to derive heterozygotes of gene-targeted animals.

Applications. Gene knockout/knockin in mice provides an effective means of investigating site-specific alterations made to the genome of a mammal. This powerful technique allows investigators to develop a mouse line with a desired modification in one or more genes and evaluate the resulting loss-of-function or gain-of-function phenotype. When coupled with Cre and inducible approaches, this service offers investigators relatively precise control over the temporal and spatial ablations or expression of targeted genes in vivo.

To request this service, click the “How to obtain services” link.

To learn how to prepare and deliver your gene-targeted ES cells, click the “Helpful information” link.

Establishment of gene-targeted ES cell clones by homologous recombination

The Facility will culture ES cells; introduce gene targeting constructs into ES cells by electroporation; select for and isolate gene-targeted ES cell clones; culture and expand isolated ES cell clones for genotyping; cryopreserve the isolated ES cell clones; and expand ES cell clones with a homologous recombination event. These services currently require the combined efforts of our Facility staff and lab members of the investigator requesting the service. At this time, the investigator’s lab must perform the genotyping for ES cell clones with a homologous recombination event (targeting). The J1 ES cells available from our Facility originate from the 129/Sv strain and have a good track record of efficient homologous recombination and germline transmission—two properties necessary to derive knockout/knockin animals.

Applications. Homologous recombination allows gene targeting at desired loci, and germline transmission enables the establishment of mouse lines.

To request this service, click the “How to obtain services” link.

To learn how to prepare your transgene constructs for injection, click the “Helpful information” link.

Forms

Request Form for Service  (for Wistar users)

Request Form for Service  (for non-Wistar users)

Archive of Submitted Materials  (All users)