Construction of severe combined immunodeficiency mice based on CRSIPR/Cas9 technology / 中国实验动物学报

Objective To knockout Rag2 and IL2rg genes and construct severe combined immunodeficiency mice based on CRISPR/Cas9 technology. Method Design and synthesis of 25 bp sgRNA were made according to the Rag2 and IL2rg sequences in Genbank. After annealing, sgRNA was cloned into pX330 vector. Recombination plasmid Rag2?sgRNA, IL2rg?sgRN and Cas9 were then transcribed into RNA, these RNA were microinjected into zygotes and the zygotes were transplanted into recipient ICR mice. F0 founders were born and mutated F0 founders mated with wild type mice to obtain F1 generation heterozygous mice. Mutated F1 mice were crossed and got F2 generation homozygous mice. Genotype and phenotype of the knockout mice were identified by sequencing, flow cytometry and xenograft model. Results Rag2?sgRNA and IL2rg?sgRNA recombination plasmids were constructed and transcribed into RNA. After microinjection and mat? ing, F0 founders were born and F2 homozygous mice were obtained. The results of sequencing showed that there were two types of genotype in IL2rg gene, 10 bp or 11 bp deletion;however, there was only one genotype in Rag2 gene, which was 8 bp deletion. Compared with wild?type BALB/c mice, the number of CD3 +, B220 + and NKp46 + cells in peripheral blood of the knockout mice was reduced significantly. After inoculation of human breast cancer cell line SKBR?2HL cells, tumor size in the xenograft mouse model was increased gradually along with time extension. Conclusion CRISPR/Cas9 is an efficient way to mutate Rag2 and IL2rg gene in mice in vivo, leading to aberrant T cells, B cells and NK cells.

Severe combined immunodeficiency mouse-psoriatic human skin xenograft model: A modern tool connecting bench to bedside.

Psoriasis is a multifactorial chronic inflammatory disease. Research into the pathogenesis of this disease is hindered by the lack of a proper animal model. Over the past two decades, many scientists were involved in the development of animal models that nearly mirror the immunopathogenesis of psoriasis. One such model, which has opened doors to the study of molecular complexities of psoriasis as well as its treatment, is the severe combined immunodeficiency (SCID) mouse-human skin chimera model. This model not only mirrors the clinical and histopathological features of psoriasis but also help in the study of cell proliferation, angiogenesis, function of T cells, neurogenic inflammation and cytokines involved in inflammatory reactions. In this article, we have reviewed the prospects and the limitations of the SCID mouse model of psoriasis.

Establishment of human leukemia model by transplanting K562 cells into SCID mice / 重庆医科大学学报

Objective: To establish the transplanted leukemia severe combined immunodeficiency(SCID) mouse model and to explore the biological action of human leukemic cells in SCID mice.Methods: 5?106～1?107 K562 cells were inoculated into the abdominal cavity or vein of SCID mice.Immunocytochemistry,RT-PCR,flow cytometry and histopathology were used to detect the leukemic cells in the peripheral blood,bone marrow,liver and spleen of SCID mice.Results: K562 cells could be transplanted into SCID mice through abdominal cavity or vein,and leukemia animal model was successfully established.3 weeks after transplantation,the leukemia cells could be found in the peripheral blood,The proliferation phase cells in the liver and spleen reached 40.11% and 41.66% respectively.In the terminal stage,the CD13+ cells accounted for 10.15% of bone marrow cells in model animal.The DQ?and ABL-BCR genes could be detected after injection of K562 cells,and 3 to 4 weeks after injection,the leukemic cells spread to the liver,spleen,kidney,bone marrow,and so on.Conclusion: The leukemia animal model could be successfully established by transplanting K562 cells into SCID mice.Positive rate of CD13 might be used as the index of tumor cells infiltration.

Study of anti-glioma of SEC in vivo and in combination with radiotherapy for glioma / 中国免疫学杂志

Objective:To observe the anti-glioma effects of lymphocyte activated by SEC in vivo and probe into therapy effects of SEC combing with operation,radiotherapy and chemotherapy.Methods:Experiment animals were SCID mice.With establishing animal model of glioma xenografted and immunity chimeric model of HuPBL-SCID,we observed the growth curve,survive time.With having operation,radiotherapy and chemotherapy,30 patients suffering from glioma were divided into control group,handle group 1 and 2,and then results were analyzed with MRI.Results:Growth curve shows:A,C and B group produced different suppress against glioma and its suppress rate were 58.5%,46.2% and 36.3%.The first death occurred at the 17th day in control group mice and all mice deaded at the 40th day.Contrasting to this,the first death occurred at the 40th day and 50% mice still were alive in group A until the day that experiment has just finished(60 day).Clinical material showed:The rate of effect(CR and PR) in control group was 40%,while the rate of effects was 50% in handle group 1 and 62.5% in handle group 2.Conclusion:SEC has extramaly potent anti-tumor activity against glioma.SEC combined with operation,radiatherapy and chemotherapy can raise treatment effect in patients with glioma.