In the past several years, our cores top priority has been keeping up with the latest developments in genome engineering methods, especially the CRIPSR (clustered regularly interspaced short palindromic repeat) technology. We can now efficiently generate knockout mice by creating small frame shift mutations or generating large deletions by cutting with two or more CRISPR constructs. We can also use single strand oligonucleotides as donors to efficiently knockin point mutations or insert small pieces of DNA, such as loxP or various tags. In the last 12 months, we have assisted more than twenty NIH laboratories in over three dozen knockout/knockin projects. We also completed four conditional knockout projects using the CRISPR method. We have also made progress in knocking in large genes using double strand donor vectors. Besides developing the new genome engineering technologies, our core is continuing to provide a variety of services using the classical mouse genetic and reproductive methodologies, including generating transgenic mice using the pronuclear microinjection method, generating knockout mice using the conventional ES cell-mediated homologous recombination, carrying out mouse in vitro fertilization, re-deriving and resurrecting mouse lines. We have also made technical advancements in using such traditional transgenic and ES cell-mediated methods. For examples, we have produced excellent chimeric mice using an ES cell line derived from the PWD mouse strain, and have inserted a large transgene into the Rosa26 locus using the so-called site-specific (TARGATT) transgenic technology. In addition, we are also continually expanding our capabilities in using immunocompromised mice to support stem cell-related research. Our services in this area have extended to beyond teratoma assays, i.e. searching for representative tissues from all three germ layers. The cell types we inject have broadened to include differentiated cells and cancer cells, and the assays have also expanded to analyzing human cell engraftment and testing the efficacy of experimental drugs.

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Project End
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Budget End
Support Year
9
Fiscal Year
2016
Total Cost
Indirect Cost
Name
U.S. National Heart Lung and Blood Inst
Department
Type
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Lin, Yongshun; Liu, Huimin; Klein, Michael et al. (2018) Efficient differentiation of cardiomyocytes and generation of calcium-sensor reporter lines from nonhuman primate iPSCs. Sci Rep 8:5907
Zhang, Yingfan; Liu, Chengyu; Adelstein, Robert S et al. (2018) Replacing nonmuscle myosin 2A with myosin 2C1 permits gastrulation but not placenta vascular development in mice. Mol Biol Cell 29:2326-2335
Jiao, Delong; Cai, Zhenyu; Choksi, Swati et al. (2018) Necroptosis of tumor cells leads to tumor necrosis and promotes tumor metastasis. Cell Res 28:868-870
Zhuang, Lenan; Jang, Younghoon; Park, Young-Kwon et al. (2018) Depletion of Nsd2-mediated histone H3K36 methylation impairs adipose tissue development and function. Nat Commun 9:1796
Xing, Shaojun; Shao, Peng; Li, Fengyin et al. (2018) Tle corepressors are differentially partitioned to instruct CD8+ T cell lineage choice and identity. J Exp Med 215:2211-2226
Liu, Tanbin; Hu, Yi; Guo, Shiyin et al. (2018) Identification and characterization of MYH9 locus for high efficient gene knock-in and stable expression in mouse embryonic stem cells. PLoS One 13:e0192641
Vizcardo, Raul; Klemen, Nicholas D; Islam, S M Rafiqul et al. (2018) Generation of Tumor Antigen-Specific iPSC-Derived Thymic Emigrants Using a 3D Thymic Culture System. Cell Rep 22:3175-3190
Deis, Jessica A; Guo, Hong; Wu, Yingjie et al. (2018) Lipocalin 2 regulates retinoic acid-induced activation of beige adipocytes. J Mol Endocrinol :
Lee, Hye Kyung; Willi, Michaela; Wang, Chaochen et al. (2017) Functional assessment of CTCF sites at cytokine-sensing mammary enhancers using CRISPR/Cas9 gene editing in mice. Nucleic Acids Res 45:4606-4618
Xu, Zhe; Xing, Shaojun; Shan, Qiang et al. (2017) Cutting Edge: ?-Catenin-Interacting Tcf1 Isoforms Are Essential for Thymocyte Survival but Dispensable for Thymic Maturation Transitions. J Immunol 198:3404-3409

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