PiggyBac-on-BAC Transgenic Mice

Our new PiggyBac-on-BAC transgenic mouse service can deliver single-copy transgenic mice with more consistent expression of your transgene. This proprietary transgenic method has the following advantages over other transgenic approaches allowing for increased reliability and consistency in expression:

(1) Integration occurs as a single copy at each integration site. By contrast, in conventional transgenics, there are typically multiple copies integrated in tandem at each integration site. The presence of multiple copies can interfere with the expression of the transgene, and may even induce silencing.

(2) The integrated fragment is defined, namely only your desired transgene sequence is integrated. No vector backbone sequence is integrated, nor is there any loss of your transgene sequence. By contrast, in conventional transgenics, the transgene may integrate along with extra vector backbone sequence, or the transgene sequence may be partially eroded.

Workflow of PiggyBac-on-BAC Transgenic Mouse Projects

Each full PiggyBac-on-BAC transgenic mouse project is divided into the following phases:

1. PiggyBac-on-BAC transgenic strategy design

• Design a PiggyBac-on-BAC transgenic strategy that meets your research needs.
• Develop a PCR genotyping strategy for PiggyBac-on-BAC transgenic founder screening.

2. PiggyBac-on-BAC modification

• Modify BACs by recombineering as needed for BAC-based transgenes (visit our BAC Modification Services page for details).
• Confirm the BAC modification by restriction digest and sequencing as needed.

3. PiggyBac-on-BAC transgenic pronuclear injection

• Inject the verified PiggyBac-BAC into the pronucleus of fertilized eggs.
• Implant injected eggs into surrogate mothers to obtain offspring.

4. PiggyBac-on-BAC transgenic mouse screening

• Genotype pups by PCR to identify those carrying the PiggyBac-on-BAC transgene.

Technical Details of PiggyBac-on-BAC Transgenic MIce 

Preparation of DNA for injection and development of a genotyping strategy

The success of PiggyBac-on-BAC transgenic production depends critically on the design and the quality of PiggyBac-BAC used in pronuclear injection. Accordingly, we employ a special protocol to design and prepare top-quality PiggyBac-BAC for our injections. BAC DNA is typically injected in unlinearized form, as this has been shown to produce similar biological outcomes as linearized BAC.

Generation of PiggyBac-on-BAC transgenic founder mice

-PiggyBac-BAC injection to obtain founders, basic service: A minimum of 200 eggs (C57BL/6) will be injected to produce live animals. This will result in an average of 2 transgenic founders, but the actual numbers could fluctuate significantly, and it is possible that no founders are obtained. If you wish to ensure the generation of founders, we strongly recommend that you use our guaranteed service option (see below).

-PiggyBac-BAC injection to obtain founders, guaranteed service: We guarantee a minimum of 2 transgenic founders (the actual number of founders obtained is typically higher).

-Genotyping pups to identify founders: Pups will be genotyped by PCR to identify PiggyBac-on-BAC transgenic founder mice following the strategy designed by Cyagen scientists.

Pricing and Turnaround Time

Guarantee

Cyagen offers the best guarantee in the industry – we will fully refund the client’s service fee if animals with the specified genotype are not generated (except for genetic modifications severely affecting viability, morbidity, or fertility). Given the complexity of biological systems, a particular genetic modification may not result in the desired phenotype. As such, Cyagen's guarantee covers the creation of animals with the specified genotype, not a particular phenotypic outcome in terms of transcription, protein/RNA function, or organismal biology.

Order Supporting Information 

Please see below for supporting information related to Cyagen animal orders.

（https://www.cyagen.com/us/en/support/order-information.html）

Inquiries and Quote Requests

Request a quote now. Alternatively, you can always email animal-service@cyagen.com or call 800-921-8930 to inquire about our services or obtain a quote for your project.