Novel Model Construct Validation

Are you working with a newly made transgenic mouse model to test therapeutics? Are you confident that the results you’re seeing are an effect of the therapeutic or could there be a genetic anomaly influencing your phenotype?

Researchers commonly initiate characterization studies on transgenic lines prior to confirming the stabilization of transgene copy number and genetics. This can be a costly error resulting in lost time, funds, and other valuable resources. It could incorrectly eliminate promising therapeutics or delay the validation of a therapeutic.

Not only can this delay publication of results, but your results may be irreproducible!

Transgenes integrate into the host genome at a random location, typically in a head to tail array of multiple copies. Newly made transgenic founders may be mosaic, and harbor multiple integrations on different chromosomes. Very high copy number transgenic founders may be vulnerable to copy number loss that can influence phenotype.

GenoTyping Center of America (GTCA) can help you avoid these pitfalls with quick and cost-effective transgenic model validation. This quality assurance validation is imperative to properly begin any research project relying on transgenic lines.

GTCA’s unique Novel Model Construct Validation Service will confirm your transgenic model to ensure your research efforts are built on a solid foundation. Our experienced team will…

verify the 5’, middle and 3’ end of your construct,
assess the relative transgene copy number and determine that it is inherited stably generation to generation without evidence of copy number alteration,
analyze your data to see if your model is exhibiting signs of multiple integrations,
formulate a management plan so that your model continues to provide reproducible results generation after generation,
provide zygosity (Hemizygous vs Homozygous) genotyping for transgenics, even when the integration site is unknown

Experimental Consequences of Working with Unstable Transgenic Lines

Multiple Integrations

Transgene integrates on multiple chromosomes and each integration segregates independently.

The Consequences

Too many carrier animals are observed in the progeny. Carrier animals may harbor one or both of the integrations, and each variant can give rise to a different phenotype.

The Fix

GTCA scans all progeny by quantitative PCR. Selective breeding of high and low copy variants is preformed until copy number is stable and Mendelian genetics are observed. once stable, sublines are phenotyped knowing that variable transgenic integration caveats have been eliminated.

Play Video

CRISPR-Cas9 Founder Identification & Line Development Planning Webinar – A short, but highly informative, overview narrated by GTCA’s Melissa Osborne.

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