Transgenic Mouse Service

You Think it up, we knock it out (or in)

As a scientist, you have enough on your plate. Not sure which type of transgenic mouse would work best for your experiment? We've summarized some of our most popular transgenic mouse services and model types below.

Don't see the exact fit for your application? Contact us for personal assistance designing the customized mouse model that is right for you.

What Researchers Say

"I've been working with iTL over the past 5 years in the production of 3 different genetically altered mice. Not only did iTL help in the design of the mice, but the entire process was transparent with the opportunity at any time along the way to discuss my questions or concerns with scientists who had significant insight into the process. The mice were delivered on time, as billed!"

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— Raghu Mirmira, MD, PhD, University of Chicago

Custom Transgenic Mice

Types of Transgenic Mice Available at ingenious

Transgenic mice, by definition, are mice that have foreign DNA added into their genome. When these models were first created it was only possible to insert new sequences randomly with no control over their final genomic location.

This random integration of DNA into the genome can, and often does, create a whole host of totally unpredictable problems, such as interrupting an existing gene or leaving the new sequence somewhere in the genome that is prone to silencing, effectively eliminating any expression of your transgene.

Customized genetically modified (transgenic) models heavily impact the scientific value of your studies, as the models are designed to specifically address your scientific questions while avoiding the unpredictable complications possible in genetically modified models.

Effective research depends on finding the most relevant model for your application. When your project calls for targeting transgenesis technology, our team of specialists offer an integrated solution for the creation of your transgenic mouse model.

With ingenious, you think it up, we knock it out (or in).

Rosa 26 Targeted Transgenic Insertions

Targeted transgenic insertions were developed to circumvent the silencing and gene interruption problems encountered with random insertion of foreign DNA into the genome. One such targeted insertion site is the Rosa26 locus, discovered by Philippe Soriano's lab in 1991.

The Rosa26 locus was identified by randomly inserting Gen-ROSAβgeo provirus (genetic material of a virus) DNA into the mouse genome. The investigators used a low multiplicity of infection (ratio of viral genomes added per cell) in order to insert a single copy of provirus DNA into the genome.

The observable effect in this case was LacZ staining afforded by the βgeo reporter gene present in the Gen-ROSAβgeo proviral DNA. Mouse embryos expressing βgeo at the Rosa26 locus (Rosa26βgeo) expressed LacZ ubiquitously in every tissue and at every developmental stage.

Discovery of the Rosa26 locus revolutionized the development of knockin transgenic mouse models by creating a genomic safe harbor (GSH) for transgenes.

Additional Transgenic Mouse Services

Sperm Cryopreservation

Sperm cryopreservation is a reliable, efficient and economical way to preserve transgenic mouse lines. Sperm are collected from two male mice and a motility test is completed for thawed sperm. This service follows thawed cryopreserved sperm through in vitro fertilization (IVF) all the way to the blastocyst stage to ensure sperm fertility.

ingenious offers many different storage plans to fit your specific needs and a cryorecovery service is available, if needed, to successfully reestablish your mouse line.

Benefits of Sperm Cryopreservation:

• Easy distribution of your transgenic line to other researchers
• Easy restoration of a specified and opportunistic pathogen free (SOPF) mouse line
• Significant financial benefit for low use strains rather than maintaining a live colony
• Limits the genetic drift of a colony

The sperm cryopreservation method is best suited to transgenic mice possessing single mutations on inbred backgrounds.

Embryo Cryopreservation

Embryo cryopreservation is an alternative method to preserve your transgenic mouse line. While the method requires more mice from your colony to achieve, the preserved embryos will maintain the correct genotype of your transgene (homozygous versus heterozygous) and the specific genetic background of the mice.

This may be advantageous when homozygous mice are required and transgenic sperm cryorecovery generates only heterozygous pups.

Embryo cryopreservation is best suited for transgenic mice with multiple mutations.

Breeding Services and Mouse Colony Management

Are you new to transgenic mouse colony management or just don't have the resources for the task? ingenious provides comprehensive mouse breeding and colony management services, whether you require a small or large cohort of homozygous or heterozygous transgenic mice and age-matched controls.

Our highly skilled team genotypes all mice and validates all mouse lines based on optimized PCR protocols and tightly controls all animal housing costs.

Based on your research requirements, we can time the delivery of age-matched experimental and control cohorts directly to your facility. We can additionally cross your transgenic mouse with a Cre (or tTA, tTS or CreERT) line, creating a conditionally expressed transgenic mouse line.

Breeding Schemes

Depending on the effects your transgene may have on mouse fertility and viability, three different breeding schemes are possible for your new transgenic mouse colony. With each breeding scheme, different proportions of heterozygous and homozygous offspring will be produced and genotyping may or may not be required, affecting your costs.

1) Heterozygous mutant (+/-) x Heterozygous mutant (+/-)

• Used when neither sex of homozygous mutant mice is viable or fertile
• Produces 25% homozygous wild type (WT), 50% heterozygous and 25% homozygous mutant mice
• All mice must be genotyped

2) Heterozygous mutant (+/-) x Homozygous mutant (-/-)

• Used when only one sex of homozygous mutant mice is viable and fertile
• Produces 50% heterozygous and 50% homozygous mutant mice
• All mice must be genotyped

3) Homozygous mutant (-/-) x Homozygous mutant (-/-)

• Used when both sexes of homozygous mice are viable and fertile
• Produces 100% homozygous mutant mice
• No need to genotype mice

Colony Management Assistance

ingenious also provides transgenic mouse colony management assistance for those laboratories unfamiliar with the technologies and procedures involved with caring for their transgenic mice. We facilitate the teaching and training of staff members on an as needed basis, as defined by the client, to ensure the optimization of all colony management procedures.

Additionally, we assist in setting up breeding schemes to match future needs and database management. ingenious uses SoftMouse software for all mouse colony management.

Genotyping Services

Genotyping your mouse colony is simple in theory, but can be difficult in practice. Establishing the correct genotypes of your mouse colony with absolute certainty is imperative for sound research studies and can avoid the costly scenario of losing months worth of work due to genotype uncertainty and inconsistent results.

At ingenious, our PCR experts minimize PCR inhibitors in extracted mouse DNA samples and optimize our PCR to ensure efficient, highly specific reactions. We test your transgenic mouse DNA under rigorously controlled conditions so you can have 100% confidence in our genotyping.

We offer genotyping to distinguish between heterozygous and homozygous WT and mutant mice and testing for multiple transgenes per mouse. Outsourcing colony genotyping to the experts at ingenious may be some of the smartest research dollars you could ever spend.

Tissue Harvesting

ingenious provides tissue harvesting services for researchers that require mouse tissues but not an entire mouse. We can harvest nearly any tissue you require, prepared to your specifications, and delivered directly to your facility.

Our organ, tissue and blood extractions are performed by highly experienced surgeons using the most stringent aseptic techniques. We routinely collect over 75 different specimens and special requests are considered.

Mouse Colony Management: Breeding Planner

Not sure whether you should house your own transgenic mouse colony or outsource the work to ingenious? We provide you the cost breakdown of maintaining a live mouse colony at the cohort size, sex and age you require with our free online Breeding Planner.

By entering the number of homozygous transgenic, heterozygous and WT males and females you need and the costs to genotype and house each mouse, you can get an accurate picture of the amount of time and money that will be required to build up your transgenic mouse colony.

Importantly, the Breeding Planner also includes detailed breeding process milestones and a breeding strategy diagram that will assist your staff in planning the breeding crosses and achieving milestones set to keep the colony on schedule.

Does it all sound like a lot of work? Contact us about outsourcing your colony breeding to the experts at ingenious.

References

1. Friedrich G1, Soriano P. 1991. Promoter traps in embryonic stem cells: a genetic screen to identify and mutate developmental genes in mice. Genes Dev 5 (9): 1513-23.

2. Zambrowicz BP, Imamoto A, Fiering S, Herzenberg LA, Kerr WG, Soriano P. 1997. Disruption of overlapping transcripts in the ROSA beta geo 26 gene trap strain leads to widespread expression of beta-galactosidase in mouse embryos and hematopoietic cells. Proc Natl Acad Sci USA 94 (8): 3789-94.

3. Kisseberth WC, Brettingen NT, Lohse JK, Sandgren EP. 1999. Ubiquitous expression of marker transgenes in mice and rats. Dev Biol 214 (1): 128-38.