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Vanderbilt Genome Editing Resource

piggyBac Transgenesis

Insertion of DNA randomly into the mouse genome is easily achieved using piggyBac transgenesis. VGER maintains stocks of piggyBac transposase and piggyBac transposons can be sourced from Addgene and VectorBuilder.com.

Benefits of piggyBac transgenesis compared to standard transgenesis approaches:

  • Single copy insertions
  • High transposition efficiency
  • Tendency to land within transcriptionally active chromatin loops
  • Transgenes are less likely to be silenced
  • Large cargo capable (up to 200 kb in the literature)

Downsides to any random DNA integration method is the challenge of mapping insertion sites and the potential position effects on transgene expression patterns and levels. VGER has developed methods to map piggyBac transposon insertions.

Full service projects include:
  • Project consultation
  • Transgene design
  • DNA quality control and injection solution preparation
  • Embryo microinjection
  • Screening of founder mice
  • Breeding of founders to confirm germline transmission
  • Assessment of copy number
  • Inverse PCR mapping
  • Assignment of official MGI allele name and number
  • Development of an automated genotyping assay in conjunction with TransnetYX
  • Support for manuscripts and grant applications

Workflow:

Advisory Meeting → Strategy Design → Reagent Procurement → Editing of Mouse Embryos → Screening of F0 Generation → Breeding and Screening of N1 Generation

Average Time to F0 Founders:

5-6 months

Price through F0 Founder Generation:

Shared resources must adjust pricing annually. Projects are by custom quote and depend on project complexity.

Genotype Guarantee:

As a shared resource, we cannot offer a money back guarantee. Based on previous projects, we can confidently state that large knock-ins that do not severely affect viability, morbidity, or fertility, that are introduced into the standard strains, C57BL/6J or C57BL/6N, will be produced within two pronuclear injection attempts, with within a single attempt.

Models Produced: 10