NCG小鼠
品系名称：NOD/ShiLtJ-Prkdc em26Cd52 Il2rg em26Cd22
基因官方名：Prkdc;Il2rg
基因全名：protein kinase DNA activated catalytic polypeptide蛋白激酶DNA活化催化多肽；interleukin 2 receptor gamma chain 白细胞介素2受体γ链
基因别名：AI326420；AU019811；DNA-PKcs；DNAPDcs；DNAPK；DNPK1；DOXNPH；HYRC1；XRCC7；dxnph；p460；
NCBI号：19090,16186
品系背景：NOD/ShiLtJ
修饰方式：Knockout (KO) CRISPR/Cas9
保存方式：活体 冷冻
微生物级别：SPF级
品系描述
NCG (NOD/ShiltJ-Prkdc em26Cd52 Il2rg em26Cd22)是使用基因编辑技术敲除了NOD/ShiltJ小鼠的Prkdc (Protein kinase, DNA activated, catalytic polypeptide)及Il2rg（Common gamma chain receptor）基因而获得的重度免疫缺陷品系。NOD/ShiltJ遗传背景使该品系具有天然免疫缺陷，如补体系统、巨噬细胞缺陷，同时，该背景Sirpa与人类CD47具有高亲和力，使NOD/ShiltJGpt比其它品系更适合人源移植物（如：肿瘤和人源细胞）的定植。Prkdc基因功能缺失导致V(D)J重组不能正常发生，造成T细胞和B细胞不能发育成熟。IL2RG是多种白细胞介素细胞因子受体的共同亚基，IL2RG失活则导致6种不同细胞因子信号通路缺失，造成NK细胞缺陷。因此，NCG是迄今为止免疫系统缺陷最为彻底的小鼠模型之一，非常适合人源肿瘤细胞移植（CDX）、人源肿瘤组织移植（PDX）、人外周血单个核细胞（PBMC）及人源造血干细胞（CD34+ HSC）移植进行免疫重建。NCG生存周期长（>89周），利于长期移植及药效学评价。
Description
Severe immune-deficient strain NCG is established by GRISPR/Cas9 technology. Prkdc(Protein kinase, DNA activated, catalytic polypeptide)and Il2rg （ Common gamma chain receptor）genes are knocked out on NOD/ShiltJGpt background.The genetic background of NOD/ShiltJGpt makes this line have natural immunodeficiency, such as complement system and macrophage defects[1]. At the same time, the Sirpa on NOD/ShiltJGpt has high affinity with human CD47, making it more suitable for colonization of human grafts (e.g. tumors and human cells) than other strains[2]. Loss of Prkdc gene leads to the inability of V(D)J recombination to occur, resulting in the inability of T cells and B cells to mature. Il2rg is a common subunit of various interleukin cytokine receptors, and the inactivation of Il2rg leads to the loss of six different cytokine signaling pathways[4], resulting in NK cell defects[3].Therefore, NCG is the most thorough mouse model of the immune-deficient to date, and is very suitable for Cell derived xenograft (CDX), Patient derived xenograft (PDX), human peripheral blood mononuclear cells (PBMC) and human hematopoietic stem cell(CD34+HSC) transplantation for immune reconstitution.The NCG has a long life cycle of >89 weeks, which is beneficial for long-term transplantation and pharmacodynamic evaluation.
Genotyping Protocol
Genetic Background：NOD
Strain Type：KO
Gene Name：Prkdc; Il2rg
1. Strategy of Genotyping
Wild type: ①PCR reaction obtains a single WT band; ②PCR reaction obtains a single WT band.
Heterozygote: ①PCR reaction obtains a WT band and a KO band; ②PCR reaction obtains a WT band.
Homozygote: ①PCR reaction obtains a single KO band; ② PCR reaction without product.
Note：1)The sizes of WT and Targeted band are shown below.
2)If the WT band is too large, it may not be possible to obtain a WT band.
2.2 Prkdc Primer Information
2.3 Il2rg Primer Information
3. Gel Image
Note: P: Positive control; B6: Wildtype control; N: Blank control (ddH2O)
4. PCR Condition
应用领域
1、人源免疫重建小鼠模型，如BLT人源化小鼠、PBMC人源
化小鼠及CD34+人源化小鼠
2、人源肿瘤细胞、肿瘤组织移植（CDX，PDX）
3、药效评价（小分子、大分子、联合用药）
4、人类癌症模型
5、干细胞研究
Application
1. Full humanized immune system reconstitution mouse model
(CD34+hematopoietic stem cell engraftment)
2. Cell derived xenograft (CDX), Patient derived xenograft (PDX)
3. Immune-oncology and efficacy test
4. Human cancer model
5. Stem cell research
验证数据 Data support
■ NCG免疫细胞分群检测图 T/B/NK cell ratio assay
取5周龄NOD（NOD/ShiltJ）、NOD-scid及NCG雌鼠脾脏细胞进行流式检测，测定其T细胞（CD3+）、B细胞（CD19+）、NK细胞（CD335）及成熟B细胞（IgM+）比例。结果显示，与NOD相比，NOD-scid几乎没有T、B细胞，NK细胞比例代偿性上升。与NOD相比，NCG几乎没有T、B、NK细胞，免疫缺陷比NOD-scid更为彻底。
Detection of T/B/NK cell ratio in NCG mice. Proportion of CD3+ (T cell), CD19+(B cell), CD335+(NK cell) and IgM+ (mature B) cells is in the splenocytes from NOD/ShiLtJGpt, NOD-scid and NCG females was analyzed by flow cytometry after stained with appropriate antibody. The result indicate there was almost no detectable T cell, B cell and NK cell in NCG mouse, confirmed that NCG is the severe immune deficient model.
■ NCG小鼠在CDX中应用 PDX in NCG
PDX model (Patient-derived xenograft model) are established through directly transplanting patient's tumor tissue into the immune-deficient mice (NCG), which retains the characteristics of most of the primary tumors in histopathology, molecular biology and gene expression. As a result, PDX model is widely used in the development of new drugs and predictive biomarkers.
■ NCG小鼠在PDX中应用
■ NCG小鼠在CDX中应用 CDX in NCG
The cell line-derived xenograft (CDX) model is a well-recognized and commonly used model for the research and development of anti-tumor drugs. The CDX model is also widely used in the early stage of in vivo study because of its user-friendly technology and excellent repeatability. Cancer cell are usually inoculated to immune-deficient mice (NCG) subcutaneously, intravenously or orthotopically. In vivo imaging system could be used to monitor tumor growth with orthotopic inoculation.
■ huPBMC-NCG
■ huHSC-NCG
■ 活体成像 In vivo imaging system
参考来源：综合参考文献报道、集萃药康、JAX、百科词条等。
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