TARGATT™ attP Mice
Make your own TARGATT™ knock-in mouse for gene overexpression, reporter gene insertion, knock-down by siRNA, humanized models and other applications.
The TARGATT™ attP mice (C57BL6) contain an integrase recognition docking site at the mHipp11 safe harbor locus (on mouse chromosome 11). These mice can be used for site-specific and stable integration of your transgene into the safe harbor locus, with high insertion efficiency and a fast turnaround.
The TARGATT™ attP mice, TARGATT™ plasmids (with choice of different promoters), and associated TARGATT™ kits enable you to make site-specific transgenic animals in your own lab. This is especially useful for transgenic animal core facilities, where many different lines of transgenic mice need to be created.
How-to Guide for TARGATT™ Transgenic Kit: Dr Ruby Chen-Tsai
Sample Health Report:
The TARGATT™ “attP” C57L6/H11 mice (CRL strain code #549) are available for order directly from Applied StemCell (ASC). Please note: As of 12/21/17, Charles River Laboratories (CRL) will not sell TARGATT™ mice. However, CRL will continue to breed, produce and ship the TARGATT™ mice from their facility for ASC.
You can place an order for TARGATT™ C57BL6/H11 mice using one of the following ways:
(1) ASC Website
(2) Inquiry form
(4) Phone: 1-866-497-4180 (US Toll Free) 1-408-773-8007, 8am- 6pm (PST, USA)
(5) Fax: 1-650-800-7179
Shipping fee for US Domestic orders starts at $85 per crate (containing 30 female mice) by ground shipping (through World Courier). Additionally, shipment by air freight is also available, starting from $400 per crate for US Domestic customers. Please inquire .
Once you place your order with ASC, the order will be transferred to CRL system. ASC will notify the customer about the estimated delivery date based on the shipping schedule. CRL will then ship the animals directly to customer’s animal facility.
You can place orders through our website , email or inquiry form at any time (24/7); or by phone (8am-6pm PST, USA). Orders will be processed and transferred to CRL between 9 am - 12 pm (noon) PST. Please be aware that it takes a minimum of three (3) days lead time for a shipment. Orders can be processed sooner if all customer information and permits are up-to-date in our system.
It can take a minimum of 3 business days for CRL to ship the mice if all information and permits are up-to-date. The processing time may take longer for orders from new customers/ facilities, depending on the time to set-up permits for CRL to ship the animals into the receiving facility.
CRL breeds and ships animals per a tight schedule. An additional cost may be incurred for choosing a specific receiving date.
The animals are house in Building 16. The Charles River shipping address (from and building number) is:
Charles River GEMS (GEMS Internal Projects)
251 Ballardvale Street Wilmington MA 01887 USA
Yes. You can contact us for information regarding age of mice, number of animals available, etc., before placing an order.
(1) Inquiry form
(3) Phone: 1-866-497-4180 (US Toll Free) 1-408-773-8007, 8am- 6pm (PST, USA)
(4) Fax: 1-650-800-7179
Please provide the following information when placing an order: (1) Age(s) of mice; (2) Number of mice; (3) Contact information; (4) Shipping address; (5) Payment Details; (6) Shipping Methods; (7) Contact information for veterinarian/ import coordinator for your animal facility; and (8) Signed Material Transfer Agreement (MTA).
You will need to purchase the (1) TARGATT™ plasmids that contain the “attP” recognition sequence, “attB” to clone your gene of interest; the (2) TARGATT™ Transgenic Kit which contains integrase mRNA and buffers for microinjection of your TARGATT™ plasmid into the TARGATT™ mouse embryo; (3) the TARGATT™ mouse genotyping kit to genotype your TARGATT™ transgenic mice. We have a number of plasmids available with different promoters and expression modules (such as LoxP-STOP-LoxP). Please visit our TARGATT™ products page for more details.
You can also watch our “How-to” guide for a brief overview on how to generate your own transgenic knock-in mouse models using the TARGATT™ technology.
- AST-3042: TARGATT™ 2 (CAG + Poly A)
- AST-3043: TARGATT™ 3 (no promoter + MCS)
- AST-3050: TARGATT™ 6.1 (CAG-L4SL-MCS-PolyA)
- AST-3047: TARGATT™ 7 (PGK-MCS-PolyA)
- AST-3048: TARGATT™ 8 (PCA-MCS-PolyA)
- AST-3051: TARGATT™ 9.1 (PCA-L4SL-MCS-PolyA)
The TARGATT™ “attP” mice were first generated using C57BL/6N embryos. After microinjection, the founder mice were backcrossed with C57Bl/6J mice for seven generations to obtain a >99% purity of C57BL/6J genetic background, as confirmed SNP genome scanning analysis.
Applied StemCell currently sells only the TARGATT™ C57BL6/H11 mouse strain, as an off-shelf product. For other TARGATT™ mouse strains, FVB/Rosa26 and C57BL6/Rosa26 strains, please inquire.
The term, TARGATT™ “attP” mice, refers to the “docking site-ready” mouse models where the attP sequence has been inserted into either the mRosa26 or mHipp11 safe harbor loci.
The term, TARGATT™ transgenic mice, refers to the transgenic mice generated using TARGATT™ technology containing your gene of interest integrated into the safe harbor locus (i.e. a knock-in mouse model).
The TARGATT™ attP mice (C57BL6) contain the “attP” docking sites recognized by PhiC31 integrase at the mHipp11 locus on mouse chromosome 11 (H11).
- The TARGATT™ attP mice are homozygous for the attP site, and are used as embryo donors for pronuclear injection of donor plasmids containing the attB site and transgene.
- When injected together with the PhiC31 integrase mRNA, the transgene becomes permanently integrated into the embryo attP site.
- The resulting animals are screened by genotyping.
These mice can be used for site-specific and stable integration of your transgene into the H11 safe harbor locus. The TARGATT™ technology affords high efficiency insertion of transgenes (up to 22kb) with a fast turnaround and stable expression of your gene of interest.
- Chen-Tsai, R. Y. (2020). Integrase-Mediated Targeted Transgenics Through Pronuclear Microinjection. In Transgenic Mouse (pp. 35-46). Humana, New York, NY.
- Chen-Tsai, R. Y. (2019). Using TARGATT™ Technology to Generate Site-Specific Transgenic Mice. In Microinjection (pp. 71-86). Humana Press, New York, NY.
Master Cell Line
- Chi, X., Zheng, Q., Jiang, R., Chen-Tsai, R. Y., & Kong, L. J. (2019). A system for site-specific integration of transgenes in mammalian cells. PLOS ONE, 14(7), e0219842.
Description of the technology
- Zhu, F., Gamboa, M., Farruggio, A. P., Hippenmeyer, S., Tasic, B., Schüle, B., … Calos, M. P. (2014). DICE, an efficient system for iterative genomic editing in human pluripotent stem cells. Nucleic Acids Research, 42(5), e34. http://doi.org/10.1093/nar/gkt1290.
- Tasic, B., Hippenmeyer, S., Wang, C., Gamboa, M., Zong, H., Chen-Tsai, Y., & Luo, L. (2011). Site-specific integrase-mediated transgenesis in mice via pronuclear injection. Proceedings of the National Academy of Sciences of the United States of America, 108(19), 7902–7907. http://doi.org/10.1073/pnas.1019507108.
Commentary, comparison with other transgenic methods
- Rossant, J., Nutter, L. M., & Gertsenstein, M. (2011). Engineering the embryo. Proceedings of the National Academy of Sciences, 108(19), 7659-7660.
Tet inducible mice generated by TARGATT™
- Fan, X., Petitt, M., Gamboa, M., Huang, M., Dhal, S., Druzin, M. L., … Nayak, N. R. (2012). Transient, Inducible, Placenta-Specific Gene Expression in Mice. Endocrinology, 153(11), 5637–5644. http://doi.org/10.1210/en.2012-1556.
Advantage of Hipp11 (H11) locus
- Hippenmeyer, S., Youn, Y. H., Moon, H. M., Miyamichi, K., Zong, H., Wynshaw-Boris, A., & Luo, L. (2010). Genetic Mosaic Dissection of Lis1 and Ndel1 in Neuronal Migration. Neuron, 68(4), 695–709. http://doi.org/10.1016/j.neuron.2010.09.027.
Applications for mice generated by TARGATT™ (and cited/published articles)
- Carlson, H. L., & Stadler, H. S. (2019). Development and functional characterization of a lncRNA‐HIT conditional loss of function allele. genesis, e23351.
- Chande, S., Ho, B., Fetene, J., & Bergwitz, C. (2019). Transgenic mouse model for conditional expression of influenza hemagglutinin-tagged human SLC20A1/PIT1. PloS one, 14(10), e0223052. doi:10.1371/journal.pone.0223052
- Hu, Q., Ye, Y., Chan, L. C., Li, Y., Liang, K., Lin, A., ... & Pan, Y. (2019). Oncogenic lncRNA downregulates cancer cell antigen presentation and intrinsic tumor suppression. Nature immunology, 1.
- Matharu, N., Rattanasopha, S., Tamura, S., Maliskova, L., Wang, Y., Bernard, A., ... & Ahituv, N. (2018). CRISPR-mediated activation of a promoter or enhancer rescues obesity caused by haploinsufficiency. Science, eaau0629.
- Barrett, R. D., Laurent, S., Mallarino, R., Pfeifer, S. P., Xu, C. C., Foll, M., ... & Hoekstra, H. E. (2018). The fitness consequences of genetic variation in wild populations of mice. bioRxiv, 383240.
- Ibrahim, L. A., Huang, J. J., Wang, S. Z., Kim, Y. J., Li, I., & Huizhong, W. (2018). Sparse Labeling and Neural Tracing in Brain Circuits by STARS Strategy: Revealing Morphological Development of Type II Spiral Ganglion Neurons. Cerebral Cortex, 1-14.
- Kumar, A., Dhar, S., Campanelli, G., Butt, N. A., Schallheim, J. M., Gomez, C. R., & Levenson, A. S. (2018). MTA 1 drives malignant progression and bone metastasis in prostate cancer. Molecular oncology.
- Jang, Y., Wang, C., Broun, A., Park, Y. K., Zhuang, L., Lee, J. E., ... & Ge, K. (2018). H3. 3K4M destabilizes enhancer epigenomic writers MLL3/4 and impairs adipose tissue development. bioRxiv, 301986. doi:https://doi.org/10.1101/301986
- Tang, Y., Kwon, H., Neel, B. A., Kasher-Meron, M., Pessin, J., Yamada, E., & Pessin, J. E. (2018). The fructose-2, 6-bisphosphatase TIGAR suppresses NF-κB signaling by directly inhibiting the linear ubiquitin assembly complex LUBAC. Journal of Biological Chemistry, jbc-RA118.
- Chen, M., Geoffroy, C. G., Meves, J. M., Narang, A., Li, Y., Nguyen, M. T., ... & Elzière, L. (2018). Leucine Zipper-Bearing Kinase Is a Critical Regulator of Astrocyte Reactivity in the Adult Mammalian CNS. Cell Reports, 22(13), 3587-3597.
- Kido, T., Sun, Z., & Lau, Y.-F. C. (2017). Aberrant activation of the human sex-determining gene in early embryonic development results in postnatal growth retardation and lethality in mice. Scientific Reports, 7, 4113. http://doi.org/10.1038/s41598-017-04117-6.
- Nouri, N., & Awatramani, R. (2017). A novel floor plate boundary defined by adjacent En1 and Dbx1 microdomains distinguishes midbrain dopamine and hypothalamic neurons. Development, 144(5), 916-927.
- Li, K., Wang, F., Cao, W. B., Lv, X. X., Hua, F., Cui, B., ... & Yu, J. M. (2017). TRIB3 Promotes APL Progression through Stabilization of the Oncoprotein PML-RARα and Inhibition of p53-Mediated Senescence. Cancer Cell, 31(5), 697-710.
- Matharu, N., Rattanasopha, S., Maliskova, L., Wang, Y., Hardin, A., Vaisse, C., & Ahituv, N. (2017). Promoter or Enhancer Activation by CRISPRa Rescues Haploinsufficiency Caused Obesity. bioRxiv, 140426.
- Jiang, T., Kindt, K., & Wu, D. K. (2017). Transcription factor Emx2 controls stereociliary bundle orientation of sensory hair cells. eLife, 6, e23661.
- Booze, M. L., Hansen, J. M., & Vitiello, P. F. (2016). A Novel Mouse Model for the Identification of Thioredoxin-1 Protein Interactions. Free Radical Biology & Medicine, 99, 533–543. http://doi.org/10.1016/j.freeradbiomed.2016.09.013.
- Feng, D., Dai, S., Liu, F., Ohtake, Y., Zhou, Z., Wang, H., ... & Hayat, U. (2016). Cre-inducible human CD59 mediates rapid cell ablation after intermedilysin administration. The Journal of clinical investigation, 126(6), 2321-2333.
- Sun, N., Yun, J., Liu, J., Malide, D., Liu, C., Rovira, I. I., … Finkel, T. (2015). Measuring in vivo mitophagy. Molecular Cell, 60(4), 685–696. http://doi.org/10.1016/j.molcel.2015.10.009.
- Devine, W. P., Wythe, J. D., George, M., Koshiba-Takeuchi, K., & Bruneau, B. G. (2014). Early patterning and specification of cardiac progenitors in gastrulating mesoderm. eLife, 3, e03848. http://doi.org/10.7554/eLife.03848.
- Fogg, P. C. M., Colloms, S., Rosser, S., Stark, M., & Smith, M. C. M. (2014). New Applications for Phage Integrases. Journal of Molecular Biology, 426(15), 2703–2716. http://doi.org/10.1016/j.jmb.2014.05.014.
- Chen-Tsai, R. Y., Jiang, R., Zhuang, L., Wu, J., Li, L., & Wu, J. (2014). Genome editing and animal models. Chinese science bulletin, 59(1), 1-6.
- Park, K.-E., Park, C.-H., Powell, A., Martin, J., Donovan, D. M., & Telugu, B. P. (2016). Targeted Gene Knockin in Porcine Somatic Cells Using CRISPR/Cas Ribonucleoproteins. International Journal of Molecular Sciences, 17(6), 810. http://doi.org/10.3390/ijms17060810.
- Guenther, C. A., Tasic, B., Luo, L., Bedell, M. A., & Kingsley, D. M. (2014). A molecular basis for classic blond hair color in Europeans. Nature Genetics, 46(7), 748–752. http://doi.org/10.1038/ng.2991.
- Villamizar, C. A. (2014). Characterization of the vascular pathology in the acta2 r258c mouse model and cerebrovascular characterization of the acta2 null mouse. UT GSBS Dissertations and These (Open Access). Paper 508 (2014)