PiggyBac Transgenic Mice
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Our new PiggyBac 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:
(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 Transgenic Mouse Projects
Each full PiggyBac transgenic mouse project is divided into the following phases:
1. PiggyBac transgenic strategy design
- Design a PiggyBac transgenic strategy that meets your research needs.
- Develop a PCR genotyping strategy for PiggyBac transgenic founder screening.
2. PiggyBac transgenic vector construction
- Construct PiggyBac plasmid-based transgenic vectors.
- Confirm the constructed PiggyBac plasmid-based transgenic vectors by restriction digest and sequencing, as needed.
3. PiggyBac transgenic pronuclear injection
- Inject the verified PiggyBac transgenic plasmid into the pronucleus of fertilized eggs.
- Implant injected eggs into surrogate mothers to obtain offspring.
4. PiggyBac transgenic mouse screening
- Genotype pups by PCR to identify those carrying the PiggyBac transgene.
◆ Technical Details of PiggyBac Transgenic Mice
Preparation of DNA for injection and development of a genotyping strategy
The success of PiggyBac transgenic production depends critically on the design and the quality of PiggyBac vector used in pronuclear injection. Accordingly, we employ a special protocol to design and prepare top-quality PiggyBac vectors for our injections. A PCR-based genotyping strategy will be developed and tested.
Generation of PiggyBac transgenic founder mice
- PiggyBac plasmid injection to obtain founders, basic service: A minimum of 400 eggs (C57BL/6) will be injected to produce live animals. This will result in an average of 3-4 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 plasmid injection to obtain founders, guaranteed service: We guarantee a minimum of 4 PiggyBac 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 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 transgenic strategy design |
Free | 1-4 days | |
| Transgenic vector construction (PiggyBac plasmid) |
Please inquire | 2-5 weeks | |
| Prepare PiggyBac DNA for injection and develop genotyping strategy | $600 | 1-2 weeks | |
| PiggyBac plasmid injection to obtain founders, basic service | C57BL/6 | $4,950 | 8-10 weeks |
| PiggyBac plasmid injection to obtain founders, guaranteed service | C57BL/6 | $6,950 | 8-16 weeks |
| Genotyping pups to identify PiggyBac transgenic founders | $350 | 1 week |
◆ 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.
◆ 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.
◆ Animal Programs
All animal work is conducted in our specific pathogen free (SPF) facilities that have been AAALAC accredited and OLAW assured. For details information, please visit our support section for Description of our Facility, Animal Health and Animal Welfare Program.
◆ 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.