PiggyBac-on-BAC Transgenic Mice

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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 tandemly integrated copies at each integration site. The presence of tandem 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). We will inject as many eggs as needed to fulfill this guarantee.

-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.

Donor Strain Information

We typically produce transgenic mice in C57BL/6 strain backgrounds, but we may be able to offer other strains to accommodate your special research needs.

Pricing and Turnaround Time

 

Service Donor egg strain Price Turnaround time

PiggyBac-on-BAC transgenic strategy design

  Free 1-4 days
PiggyBac-BAC modification   Please inquire 2-5 weeks
Prepare PiggyBac BAC for injection and develop genotyping strategy   $450 1-2 weeks
PiggyBac-BAC injection to obtain founders, basic service C57BL/6 $5,950 10-14 weeks
PiggyBac-BAC injection to obtain founders, guaranteed service C57BL/6 $8,950 10-18 weeks
Genotyping pups to identify PiggyBac-on-BAC transgenic founders   $450 1 week
Note: For PiggyBac-on-BAC transgenic mouse services not listed above, please inquire about availability and pricing. The turnaround time above does not include the time for obtaining host institutions' approval for mouse importation, nor transit time during shipping.
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, mobidity, 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.

◆ Inquiries and Quote Requests

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

Price Matching

If you find another commercial service provider that offers better pricing than ours, we will match the price plus an additional 5% off.

◆ Payments

Standard payment terms include a 50% upfront payment before the project, and the remaining 50% plus shipping charge paid after completion of the project. If you need us to design your transgenic strategy, we will provide this service for free irrespective of whether you end up choosing us for your project.

Bulk Discount

We offer up to a 10% bulk discount for large orders. Large orders are defined as 5 or more projects from the same institution. If you bundle your orders with those of your colleagues, you can all qualify for the bulk discount.

Shipping

Products are shipped from our Santa Clara, California facility. For mouse shipments, the shipping charge includes courier cost plus a $100/crate handling fee. DNA constructs or fixed/stained embryos are shipped at room temperature, and the charge includes courier cost plus a $10 handling fee. We typically use World Courier to ship live mice and FedEx for other shipments. 

◆ Customer References

Please click here to view a map of customers who have used Cyagen before worldwide.

◆ Citations

Please click here for a list of publications that have cited Cyagen.