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Time-dependent KO rats
Rat model generation

Time-dependent KO rats

Use an inducible conditional Knockout rat to age-dependently inactivate your gene, and to enable studies at defined development stages or on age-related diseases.

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A time-dependent conditional Knockout rat defines an inducible animal model in which a gene of interest is "floxed" thus temporally controllable at a given time-point in embryonic, post-natal or adult animals.

After an additional breeding step with a Cre-inducible deleter rat line, the conditional Knockout is temporally triggered by external inducer-agents, most often small molecules such as tamoxifen or tetracycline.

Applications

For academic research:

  • Comparative study of gene function (onset vs. offset)
  • Mimic a pathology with late onset
  • Studying embryonic lethal conditions

For bio-pharmaceutical research & development:

  • Target validation studies
  • Off-target effects studies
  • Mimicking 100% antagonism
  • Compensatory phenotype rescue (compounds testing)

Strengths of time-dependent Knockout rat models

  • Very flexible: easy switch to study time-dependent KOs in different tissues
  • High physiological relevancy of the scientific data obtained from the model

Limitations of time-dependent Knockout rat models

  • Very low availability of different Cre-inducible deleter rat lines may require the creation of a new Cre-inducible rat with an adequate genetic background
    → New Cre-inducible rat lines can be produced by genOway in parallel to the conditional rat model
  • Expression levels depend on the dose of the agent administered
  • Differential penetration of trigger compound into tissue
  • Adding the loxP sites risks modification or disruption of splicing regulation (ESS ESE); possible impact on overlapping and neighboring genes
    → Need careful analysis of the placement of loxP sites
  • Tetracycline-inducible systems can leak
    → Temporal control can be achieved by using a fusion between Cre and a mutated form of the ligand-binding domain of the estrogen receptor, which only binds tamoxifen (ERt and ERt2). This inactive Cre-ERt2 fusion is activated upon tamoxifen (or 4-hydroxytamoxifen) administration.

Validation

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