PrimeCKO™ Conditional Knockout Mouse Models

Applied StemCell can generate a custom conditional knockout mouse model specific to your needs using CRISPR/Cas9 or other suitable gene editing techniques to insert the 5’ and 3’ flanking LoxP sequences into the endogenous locus of your gene of interest. These conditional knockout mice can be bred with thousands of commercially available Cre-deleter mice or custom engineered Cre-expressing mice to generate your mouse model with precise control over where and when your gene is knocked out.

  • Fast (as little as 6 months) and affordable (very low price)
  • Highly efficient CRISPR protocols and validated gRNA for precise modifications
  • Animal IP belongs to customers
  • Custom Cre- mouse generation available
  • Founder guaranteed
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Technical Details

Site-specific gene editing technologies have paved the way for novel genetically engineered mouse models with complex gene modifications that accurately represent human genetics and disease pathology. One such mouse models is the Conditional Knockout mouse model where a gene knockout can be limited to a particular tissue or a specific developmental time of the animal. This allows for better control of the gene of interest and such conditional knockout mice overcome limitations posed by constitutive knockout models such as embryonic lethality, undesired phenotypes and compensatory mechanisms. One of the most used conditional knockout method is the Cre- LoxP system, in which a gene of interest (targeted exons) is flanked by two 34-bp LoxP sequences (also called floxed alleles) at the desired genomic locus (Figure 1). The LoxP sites serve as a target for the Cre-recombinase, which catalyzes deletion of the floxed (Figure 2) exon(s).

Knockout Mouse Models Schematic Diagram

Figure 1. The schematic describes the first stage in developing a conditional knockout mouse model using CRISPR to generate a floxed (LoxP flanked exon) mouse. A single stranded donor DNA (ssDNA) is used for delivering the floxed targeting exons to replace the wild type form. The donor contains two LoxP sequences flanking the targeted exon(s) along with 5' and 3' homologous arms for directing a site-specific homology directed repair. The donor ssDNA is delivered along with Cas9 (mRNA or protein) and validated gRNAs via microinjection.

Knockout Mouse Model Crossbreeding the CKO Mouse with a Cre-recombinase Expressing Mouse

Figure 2.  Crossbreeding the CKO mouse with a Cre-recombinase expressing mouse. The Cre expression is driven by a promoter of choice: a tissue specific or ubiquitous promoter. The expressed Cre recombinase deletes the floxed exon(s) in a tissue specific manner thereby causing a frameshift in downstream sequence.

Applied StemCell has >11 years’ experience in mouse model engineering and is one of the earliest service providers of CRISPR/Cas9 technology. We can generate your CRISPR conditional knockout mouse model with full-guarantee

Learn more about our different mouse model generation services.

CRISPR/ Cas9 Conditional Knockout Mouse Service

Transgenic Mouse Model Service using homologous recombination and BAC

Publications

CRISPR Technology

CRISPR Knock-in H11 Locus in Pigs

  • Ruan, J., Li, H., Xu, K., Wu, T., Wei, J., Zhou, R., ... & Chen-Tsai, R. Y. (2015). Highly efficient CRISPR/Cas9-mediated transgene knockin at the H11 locus in pigs. Scientific reports5, 14253.

 Knock-in, Knockout, Conditional Knock-out

  • Dumesic, P. A., Egan, D. F., Gut, P., Tran, M. T., Parisi, A., Chatterjee, N., ... & Dou, F. (2019). An Evolutionarily Conserved uORF Regulates PGC1α and Oxidative Metabolism in Mice, Flies, and Bluefin Tuna. Cell metabolism.
  • Liang, T., Zhang, H., Xu, Q., Wang, S., Qin, C., & Lu, Y. (2019). Mutant Dentin Sialophosphoprotein Causes Dentinogenesis Imperfecta. Journal of dental research, 0022034519854029.
  • Qian, W., Miner, C. A., Ingle, H., Platt, D. J., Baldridge, M. T., & Miner, J. J. (2019). A human STAT1 gain-of-function mutation impairs CD8+ T cell responses against gammaherpesvirus-68. Journal of virology, JVI-00307.
  • Kweon, S. M., Chen, Y., Moon, E., Kvederaviciutė, K., Klimasauskas, S., & Feldman, D. E. (2019). An Adversarial DNA N6-Methyladenine-Sensor Network Preserves Polycomb Silencing. Molecular Cellhttps://doi.org/10.1016/j.molcel.2019.03.018
  • Deng, F., He, S., Cui, S., Shi, Y., Tan, Y., Li, Z., ... & Peng, L. (2018). A Molecular Targeted Immunotherapeutic Strategy for Ulcerative Colitis via Dual-Targeting Nanoparticles Delivering miR-146b to Intestinal Macrophages. Journal of Crohn's and Colitis.
  • Jo, S., Fonseca, T. L., Bocco, B. M. D. C., Fernandes, G. W., McAninch, E. A., Bolin, A. P., ... & Németh, D. (2018). Type 2 deiodinase polymorphism causes ER stress and hypothyroidism in the brain. The Journal of Clinical Investigation.
  • Langston, R. G., Rudenko, I. N., Kumaran, R., Hauser, D. N., Kaganovich, A., Ponce, L. B., ... & Beilina, A. (2018). Differences in Stability, Activity and Mutation Effects Between Human and Mouse Leucine-Rich Repeat Kinase 2. Neurochemical research, 1-14.
  • Amara, N., Tholen, M., & Bogyo, M. (2018). Chemical tools for selective activity profiling of endogenously expressed MMP-14 in multicellular models. ACS Chemical Biology. doi: 10.1021/acschembio.8b00562.
  • Allocca, S., Ciano, M., Ciardulli, M. C., D’Ambrosio, C., Scaloni, A., Sarnataro, D., ... & Bonatti, S. (2018). An αB-Crystallin Peptide Rescues Compartmentalization and Trafficking Response to Cu Overload of ATP7B-H1069Q, the Most Frequent Cause of Wilson Disease in the Caucasian Population. International journal of molecular sciences19(7).
  • Peng, L., Zhang, H., Hao, Y., Xu, F., Yang, J., Zhang, R., ... & Chen, C. (2016). Reprogramming macrophage orientation by microRNA 146b targeting transcription factor IRF5. EBioMedicine, 14, 83-96.
  • Hu, J. K., Crampton, J. C., Locci, M., & Crotty, S. (2016). CRISPR-mediated Slamf1Δ/Δ Slamf5Δ/Δ Slamf6Δ/Δ triple gene disruption reveals NKT cell defects but not T follicular helper cell defects. PloS one, 11(5), e0156074.
  • Besschetnova, T. Y., Ichimura, T., Katebi, N., Croix, B. S., Bonventre, J. V., & Olsen, B. R. (2015). Regulatory mechanisms of anthrax toxin receptor 1-dependent vascular and connective tissue homeostasis. Matrix Biology42, 56-73.
  • McKenzie, C. W., Craige, B., Kroeger, T. V., Finn, R., Wyatt, T. A., Sisson, J. H., ... & Lee, L. (2015). CFAP54 is required for proper ciliary motility and assembly of the central pair apparatus in mice. Molecular biology of the cell26(18), 3140-3149.
  • Bishop, K. A., Harrington, A., Kouranova, E., Weinstein, E. J., Rosen, C. J., Cui, X., & Liaw, L. (2016). CRISPR/Cas9-mediated insertion of loxP sites in the mouse Dock7 gene provides an effective alternative to use of targeted embryonic stem cells. G3: Genes, Genomes, Genetics6(7), 2051-2061.

Homologous Recombination Conditional Knockout Mouse (cited/published articles)

  • Geraets, R. D. (2019). Neuronal Ceroid Lipfuscinosis: A Tailored Animal Model of CLN2 Disease and Evaluation of Select Personalized Therapies (Doctoral dissertation, ProQuest Dissertations Publishing).
  • Zhao, M., Tao, F., Venkatraman, A., Li, Z., Smith, S. E., Unruh, J., ... & Marshall, H. (2019). N-Cadherin-Expressing Bone and Marrow Stromal Progenitor Cells Maintain Reserve Hematopoietic Stem Cells. Cell reports26(3), 652-669.
  • Li, C., Zheng, Z., Ha, P., Chen, X., Jiang, W., Sun, S., ... & Chen, E. C. (2018). Neurexin Superfamily Cell Membrane Receptor Contactin‐Associated Protein Like‐4 (Cntnap4) is Involved in Neural EGFL Like 1 (Nell‐1)‐responsive Osteogenesis. Journal of Bone and Mineral Research https://doi.org/10.1002/jbmr.3524.
  • Geraets, R. D., Langin, L. M., Cain, J. T., Parker, C. M., Beraldi, R., Kovacs, A. D., ... & Pearce, D. A. (2017). A tailored mouse model of CLN2 disease: A nonsense mutant for testing personalized therapies. PloS one, 12(5), e0176526
  • Miller, J. N., Kovács, A. D., & Pearce, D. A. (2015). The novel Cln1R151Xmouse model of infantile neuronal ceroid lipofuscinosis (INCL) for testing nonsense suppression therapyHuman Molecular Genetics24(1), 185–196http://doi.org/10.1093/hmg/ddu428.

For more journal references, please visit our comprehensive list of citations and reference publications.

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