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TurboKnockout® Gene Targeting Mice

Locate My Expert
knockout mice | knockout mouse service | model - TurboKnockout CKO Mice

Get Your Conditional Knockout & Knockin Mice in as Fast as 6 Months!

Our new TurboKnockout® service can provide you with conditional knockout, reporter knockin and humanization mouse models as fast as 6-8 months. This proprietary gene targeting method is enabled by two innovations:

(1) A super competent ES cell line that generates 100% ESC-derived founder mice rather than chimeras, eliminating the need to screen for germline transmission.

(2) A selection cassette on the targeting construct capable of removing itself after ES cell targeting without the need to breed to Flp deleter mice.

These innovations eliminate two generations of breeding, shortening production time by 4-6 months as compared to industry standard. For conditional knockouts, it may be possible to breed founders directly to your tissue-specific Cre mice.

All projects are backed by our full money-back guarantee – no animals, no charge!

◆ Workflow of TurboKnockout® Gene Targeting Mouse Services

Each full gene targeting project is split into the following phases:

knockout mouse model - Cyagen Mouse

 

1. TurboKnockout® gene targeting strategy design

  • Analyze relevant information for your target gene, including gene structure, neighboring genes at the genomic locus, known targeted animal models, etc.
  • Develop a gene targeting strategy, including the method for screening targeted ES cells by PCR and confirmation by Southern blot.
  • Propose the strategy to you for review and approval.

 

2. TurboKnockout® targeting vector construction

  • Clone relevant DNA fragments corresponding to homology arms into the targeting vector.
  • Confirm construction of the final targeting vector by restriction digest and sequencing.

 

3. Targeting in ES cells

  • Electroporate targeting vector into our super competent TurboKnockout® ES cells, followed by appropriate drug selection and isolation of drug-resistant clones. Our ES cells are derived from the C57BL/6 strain.
  • Screen for targeted ES cell clones by PCR.
  • Expand up to six PCR-positive clones, which are further confirmed by Southern blot.
  • Karyotype ES cells to ensure correct chromosome number.

 

4. TurboKnockout® mouse production

  • Introduce targeted ES cells into host embryos, followed by transfer into surrogate mothers. The resulting TurboKnockout® heterozygous mutants are 100% derived from the injected ES cells. We will aim to generate a minimum of 3 mice.
  • Genotype founders by PCR to confirm their mutant status.
  • Optional: Breed to F1.
  • Delivery of mice to the customer.

◆ Pricing and Turnaround Time
Service Price Turnaround time
TurboKnockout® targeting strategy design Free 1-4 days
Construction of TurboKnockout® gene targeting vector $8,950 8-14 weeks
Gene targeting in ES cells $12,950 8-14 weeks
Karyotype ES cells to ensure correct chromosome number $350/clone 1-2 weeks
TurboKnockout® mouse production $8,950
(we will inject as many ES cell clones as needed to fulfill our guarantee)
8-12 weeks
Optional: Breeding founders to obtain F1 Please inquire*

* You can choose to receive F0 founder animals and perform their own breeding to F1 mice. To ensure the successful generation of F1 mice and germline transmission, breeding of F0 animals require adherence to Turboknockout® breeding protocol, which will be supplied to you. You can also choose to have Cyagen perform F1 breeding, which adds 1-3 months to the projected turnaround time. You can choose to breed F0 founders to wildtype mice or to other stains such as Cre deleter strains of your choice; additional service fees may apply.

 

Note: For special knockout/knockin 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 and 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 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 begins, and the remaining 50% plus shipping paid after completion of the project. If you need us to design your targeting strategy, we will provide this service for free irrespective of whether you end up choosing us for your project (other companies typically charge a few thousand dollars for this service).

◆ 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. Cells are shipped on dry ice, and the charge includes courier cost plus a $45 handling fee. DNA constructs are shipped in E. coli 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 customer who have used Cyagen before worldwide. 

◆ Citations

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

◆ Case studies on our TurboKnockout® Gene Targeting Mice

Case 1

Sensing of viral and endogenous RNA by ZBP1/DAI induces necroptosis.
The EMBO Journal. 36:2529 (2017)
Jonathan Maelfait

Abstract

Nucleic acids are potent triggers for innate immunity. Double‐stranded DNA and RNAadopt different helical conformations, including the unusual Z‐conformation. Z‐DNA/RNA is recognised by Z‐binding domains (ZBDs), which are present in proteins implicated in antiviral immunity. These include ZBP1 (also known as DAI or DLM‐1), which induces necroptosis, an inflammatory form of cell death. Using reconstitution and knock‐in models, we report that mutation of key amino acids involved in Z‐DNA/RNA binding in ZBP1's ZBDs prevented necroptosis upon infection with mouse cytomegalovirus. Induction of cell death was cell autonomous and required RNAsynthesis but not viral DNA replication. Accordingly, ZBP1 directly bound to RNA via its ZBDs. Intact ZBP1‐ZBDs were also required for necroptosis triggered by ectopic expression of ZBP1 and caspase blockade, and ZBP1 cross‐linked to endogenous RNA. These observations show that Z‐RNA may constitute a molecular pattern that induces inflammatory cell death upon sensing by ZBP1.

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