PiggyBac Transgenic Mouse Embryos
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Our new PiggyBac transgenic mouse embryo service can deliver to you single-copy transgenic embryos with more consistent expression of your transgene. This proprietary transgenic method has three major 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.
(3) Due to the above reasons, transgene expression using our PiggyBac transgenic method is much more reliable and consistent as compared to conventional transgenics. This is confirmed by our data.
◆ Workflow of PiggyBac Transgenic Mouse Embryo 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 embryo screening
- Embryos harvested at the desired embryonic stage will be genotyped by PCR and lacZ stained to identify the ones carrying the desired PiggyBac transgene.
◆ Technical Details of PiggyBac Transgenic Mouse Embryos
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 the 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 embryos and staining of embryos for lacZ expression
- Plasmid injection to obtain PiggyBac mouse embryos, basic service: A minimum of 100-150 eggs (C57BL/6) will be injected. Embryos will be retrieved from surrogate mothers at the stage you specify (typically within the E10.5-E16.5 window) for subsequent processing. This will result in an average of 3-5 transgenic embryos, but the actual numbers could fluctuate significantly, and it is possible that no transgenic embryos are obtained. If you wish to ensure the generation of PiggyBac transgenic embryos, we strongly recommend that you use our guaranteed service option (see below).
- Plasmid injection to obtain PiggyBac mouse embryos, guaranteed service: We guarantee a minimum of 6 PiggyBac transgenic embryos for plasmid injection (the actual number of transgenic embryos obtained is typically higher).
- Genotyping and lacZ staining of PiggyBac embryos: Embryos will be genotyped by PCR. All embryos, regardless of genotype, will also be fixed and stained for lacZ expression.
◆ Donor Strain Information
We typically produce transgenic mouse embryos in C57BL/6 strain backgrounds, but we can use other strains per your request.
◆ 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 embryos, basic service | C57BL/6 | $3,750 | 5-7 weeks |
| PiggyBac plasmid injection to obtain embryos, guaranteed service | C57BL/6 | $4,250 | 5-10 weeks |
| Genotyping and lacZ staining of PiggyBac transgenic mouse embryo | $350 | 1 week |
Note: For PiggyBac transgenic mouse embryo services not listed above, please inquire about availability and pricing. The turnaround time above does not include the time for obtaining host institution’s 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, 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
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
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◆ Citations
Please click here for a list of publications that have cited Cyagen.