Standard Conditional Knockouts

Designed with you in mind

A standard conditional knockout is a versatile tool for studying your gene of interest – there are as many possible patterns of knockout as there are Cre lines. With hundreds of our conditional knockout models published, our experts will evaluate the structure of your gene and help to determine the best design for your research goals and aspirations.

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“We are actively engaged in the production of a number of conditional mouse models with iTL. This collaboration has been simple on my end, just providing the gene accession numbers of each gene, and iTL recommending the strategies for each gene. The full range of mouse knockout services matches my needs well. I find iTL’s service uniquely useful for my situation of needing different models for my research in a quick and efficient manner.”– Mehboob Hussain, MD University of Michigan Health

Standard Conditional Knockouts

Traditional Conditional Knockouts

The schematic below illustrates the targeting strategy of a conditional knockout mouse model in which exon 2 was flanked with loxP sites, and the Neo cassette was introduced into the intron downstream of exon 2. Upon FLP and Cre recombination, exon 2 is eliminated, and a loxP/FRT footprint remains.

The deletion of exon 2 results in a frame shift and early stop codons. The mRNA produced from this disrupted gene is targeted by the nonsense-mediated decay pathway, generally resulting in no protein being produced.

General Strategy Considerations

A specific target sequence within a gene of interest is flanked by loxP sites to facilitate Cre recombinase-mediated excision, for tissue specific or temporally controlled gene inactivation. The following general considerations apply for conditional knockout mouse models and are evaluated by our experts for each conditional knockout model generated at ingenious.

Preservation of wildtype expression before Cre recombination

  • Avoid insertion of exogenous sequences into regulatory regions.
  • Avoid disruption of promoter elements.
  • Avoid disruption of splice sites.
  • Insert genetic elements into large introns.

Preservation of wildtype expression before Cre recombination

  • Target the earliest exon that will result in a frameshift and premature stop codons upon deletion
  • Target an important domain.
  • Delete as much coding sequence as possible.
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