Reproductive Biology and Endocrinology
| Autor: | Dal-Young Ji, Sun-Mi Ahn, Sherrie Clark-Deener, Kiho Lee, Caitlin Ray, Eun-Jin Lee, Bumrae Cho, Jin Seok Lee, Yun-Jin Yun, Kwang-Wook Park, Jung-Taek Kang, Nathaniel Jue, Junghyun Ryu |
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| Přispěvatelé: | Animal and Poultry Sciences, Large Animal Clinical Sciences |
| Rok vydání: | 2016 |
| Předmět: |
0301 basic medicine
lcsh:QH471-489 Swine Xenotransplantation medicine.medical_treatment Spleen Biology lcsh:Gynecology and obstetrics Gene Knockout Techniques 03 medical and health sciences 0302 clinical medicine Endocrinology medicine Animals lcsh:Reproduction CRISPR CRISPR/Cas9 Alleles lcsh:RG1-991 Swine Diseases Severe combined immunodeficiency Knockout pigs Cas9 Research Somatic cell nuclear transfer (SCNT) Obstetrics and Gynecology Embryo medicine.disease IL2RG Virology Molecular biology Transplantation 030104 developmental biology medicine.anatomical_structure Reproductive Medicine 030220 oncology & carcinogenesis Models Animal Somatic cell nuclear transfer Female Severe Combined Immunodeficiency Genetic Engineering Interleukin Receptor Common gamma Subunit Severe Combined Immunodeficiency (SCID) Developmental Biology |
| Zdroj: | Reproductive Biology and Endocrinology, Vol 14, Iss 1, Pp 1-9 (2016) Reproductive Biology and Endocrinology : RB&E |
| ISSN: | 1477-7827 |
| Popis: | s Background Pigs with SCID can be a useful model in regenerative medicine, xenotransplantation, and cancer cell transplantation studies. Utilizing genome editing technologies such as CRISPR/Cas9 system allows us to generate genetically engineered pigs at a higher efficiency. In this study, we report generation and phenotypic characterization of IL2RG knockout female pigs produced through combination of CRISPR/Cas9 system and SCNT. As expected, pigs lacking IL2RG presented SCID phenotype. Methods First, specific CRISPR/Cas9 systems targeting IL2RG were introduced into developing pig embryos then the embryos were transferred into surrogates. A total of six fetuses were obtained from the embryo transfer and fetal fibroblast cell lines were established. Then IL2RG knockout female cells carrying biallelic genetic modification were used as donor cells for SCNT, followed by embryo transfer. Results Three live cloned female piglets carrying biallelic mutations in IL2RG were produced. All cloned piglets completely lacked thymus and they had a significantly reduced level of mature T, B and NK cells in their blood and spleen. Conclusions Here, we generated IL2RG knockout female pigs showing phenotypic characterization of SCID. This IL2RG knockout female pigs will be a promising model for biomedical and translational research. Electronic supplementary material The online version of this article (doi:10.1186/s12958-016-0206-5) contains supplementary material, which is available to authorized users. |
| Databáze: | OpenAIRE |
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