Conditional knockout (cKO) models are often referred to as ‘floxed’ models due to the use of Cre/lox system in their development. The Cre-Lox recombination system enables the generation of tissue-specific or inducible knockouts with a high level of control over the spatial and temporal expression of genes, and greatly improves the efficiency and specificity in generating conditional mouse.
Here is the workflow of Cre-Lox system:
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>> What applications are conditional knockout mice used for?
Compared with constitutively expressed gene mutations (i.e., knockout), conditional models provide greater temporal and regional control of gene expression - often serving as a more accurate model for studying human diseases. Researchers use cKO mouse models to bypass limitations of constitutive knockout strategies, such as embryonic lethality, modified animal physiology, compensation mechanisms, and more.
- ● Gene Function Studies: Inducible and/or tissue-specific gene expression – knockout, over-expression, etc.
- ● Target Discovery and Confirmation: cKO models solve the problem of embryonic lethality seen in KO models, enabling target confirmation in adult organisms.
- ● Verifying Compound Specificity: cKO models allow precise control of gene expression for a functional protein target, providing an in vivo system to evaluate potential off-target effects of a drug.
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>> What are methods used to create conditional knockout mice?
Cyagen provides custom cKO mouse model generation services - using state-of-the-art CRISPR, transgenic, and embryonic stem cell (ESC) technologies, such as our proprietary TurboKnockout® gene targeting - that can address the most complex project requirements.
Technology TurboKnockout® Gene Targeting CRISPR/Cas9 Gene Editing Approach Homologous recombination in ESC by
our proprietary TurboKnockout® technologyCRISPR/Cas9 nuclease mediated gene targeting
by pronuclear injectionConditional Knockout (cKO) Single target: ~7 kb
Double target: ~100 kbDonor vector: ~7 kb
ssDNA: ~100 kbModel Complexity Complex Convenient Price Moderate Economical Turnaround Time 6-8 months 5-7 months Off-target Risk No Yes Species Limitation Mice only No Donor backgrounds Mouse strains: C57BL/6, BALB/c Mouse strains: C57BL/6, FVB Rat strains:
Sprague-Dawley (SD), Long EvansIntellectual Property (IP) No risk of patent infringement Ongoing patent litigation risks Complex Genetic Modification Yes N/A
| Research Applications | Gene Target(s) | Journal Information |
|---|---|---|
| Immunology, Gut | Ddx58 |
Commensal viruses maintain intestinal intraepithelial lymphocytes via
noncanonical RIG-I signaling. Nature Immunology PMID: 31636462 |
| Cell Biology, Cardiovascular | SAHH(Ahcy) |
Inhibition of S-Adenosyl homocysteine Hydrolase Induces Endothelial
Dysfunction via Epigenetic Regulation of p66shc-Mediated Oxidative Stress Pathway Circulation DOI: 10.1161/circulationaha.118.036336 |
| Oncology | Trib3 |
TRIB3 Promotes APL Progression through Stabilization of the Oncoprotein
PML-RARa and Inhibition of p53-Mediated Senescence Cancer Cell DOI: 10.1016/j.ccell.2017.04.006 |
- Case Studies
- Testimonials
"We generated a mouse knockout model using Cyagen’s TurboKnockout® technology. Cyagen’s team was on time with milestones, provided constant updates on the project and delivered animals as promised. It was such a pleasure working with Cyagen that we decided to outsource the production of other animal models to them."
———— Maximiliano D’Angelo, Sanford Burnham Prebys Medical Discovery Institute
"Using Cyagen’s CRISPR-generated constitutive knockout mice has greatly facilitated our understanding the biology in IL-1 family of 11 cytokines or their respective receptors. We were able to have the IL-38 deficient mice and the IL-1R9 deficient mice in record time. The service provided by Cyagen is highly recommended."
———— Charles A. Dinarello, University of Colorado Denver - Anschutz Medical Campus
"Our lab has been using Cyagen’s transient transgenic mouse embryo LacZ service to test candidate sequences for enhancer activity for over five years (over 60 different sequences tested with them thus far). Their cost, turnaround time, submission process and customer service has been great, making this assay easy and routine for our lab."
———— Nadav Ahituv, University of California, San Francisco
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Technical BulletinApplications of Conditional Knockout Mice in Human Disease ResearchDiscover More >>
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>> Ready-to-use Cyagen Knockout Catalog Models
- ● Over 16,000 ready-to-use knockout (KO)/conditional knockout (cKO) mouse models
- ● No risk of failure: Our catalog features gene knockout mice that have been strictly analyzed to ensure 100% positive knockout.
- ● 100% pure B6 background eliminates need for backcrossing – most repositories have mixed backgrounds
- ● AAALAC-Accredited and OLAW Assured
- ● Delivered as Fast as 2 Weeks
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>> Custom Knockout Mouse Model Generation Services
- ● Rapid turnaround time
- ● Cost effective - low cost, fully guaranteed models
- ● "One-stop shopping" experience for customers
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>> Mouse and Rat Breeding
Cyagen’s breeding service provides reliable, economical, and efficient breeding colonies of mice/rats to support your research needs; allowing you to focus more on your research goals and less on animal care, breeding and colony management issues.
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>> Cryopreservation
Cyagen can back up your live animal colony by our embryo and sperm cryopreservation services, which can not only protect your valuable strains but also reduce your cost in the long run. We also can produce live animals for you in eight to twelve weeks once your cryopreserved stock is preserved at Cyagen.
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>> Phenotype Analysis Services
Our phenotyping services include surgical model construction alongside analytics capabilities for blood, tissue histology and morphology, pathology, genomic and proteomic expression, behavioral and metabolic analysis, and more.
Learn More >>Please contact us if you need additional information.
