Rag1-KO(Rag1-EGFP)

Nomenclature

B6;129S-Rag1tm1(loxP-EGFP-PolyA-loxP-Neo-loxP)Smoc

Cat. NO.

NM-KI-00069

Strain State

Repository Live

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Gene Summary

Gene Symbol
Rag1

Model Description

A loxP-EGFP-PolyA-loxP-Neo-loxP expression cassette was knocked into the Rag1 gene start codon site. As a Rag1 knockout mouse model, this stain can be used in subcutaneous inoculation of liver cancer tissues and tumor cells. Tumors can esaily form and grow. The amount of T and B lymphocytes in peripheral blood of mice was extremely low tested by FACS, which was comparable to or lower than that of Nude mice, and there was a significant difference compared with wild type mice. The pathological sections of HE staining of tumor tissues showed that the tumor sections of Rag1 KO mice and Nude mice were similar. This strain has the potential to replace Nude, NOD-SCID mice as a tumor-bearing mouse model.
Research Application:Immunodeficiency,tumor-bearing model

Validation Data

构建策略

rag1.jpg

图1 Rag1构建策略图。

Rag1、Rag2基因敲除小鼠表达验证

Rag1-Rag2-1.png

图2 4种小鼠外周血功能T、B淋巴细胞含量流式细胞术检测结果。

通过流式细胞术对野生型C57BL/6J小鼠、NOD-SCID小鼠、Rag1-/-小鼠和Rag2-/-小鼠外周血中B细胞和T细胞的检测结果表明:Rag1-/-小鼠和Rag2-/-小鼠中的T细胞和B细胞含量较野生型小鼠有显著性降低(P<0.0001,图1A),T细胞和B细胞含量分别约为野生型的5%和25%;较NOD-SCID小鼠虽然均值略有增高,但无显著性差异(P>0.05,图1B)。

rag1-rag2-外周血.png

图3. Rag1-KO和Rag2-KO小鼠外周血中T 、B细胞缺失。

取Rag1-KO和Rag2-KO小鼠小鼠的外周血,通过FACS对其中T 、B和NK细胞的组成进行分析(A)并进行统计对比(B)。

rag1-rag2-脾脏.png

图4. Rag1-KO和Rag2-KO小鼠脾脏中T 、B细胞缺失。

取Rag1-KO和Rag2-KO小鼠小鼠的脾脏细胞,通过FACS对其中T 、B和NK细胞的组成进行分析(A)并进行统计对比(B)。

rag1-rag2-胸腺.png

图5. Rag1-KO和Rag2-KO小鼠胸腺中T 、B细胞缺失。

取Rag1-KO和Rag2-KO小鼠小鼠的胸腺细胞,通过FACS对其中T 、B和NK细胞的组成进行分析(A)并进行统计对比(B)。

Rag-2-KO.png

图6. Rag2敲除小鼠胸腺和脾脏中EGFP检测。

结果显示Rag2 KO小鼠胸腺和脾脏中有EGFP阳性的细胞,并且EGFP阳性细胞均为CD4阴性和CD8阴性细胞。

Rag1、Rag2基因敲除小鼠的成瘤实验

Rag1-Rag2.png

图7 不同背景小鼠接种A549肿瘤细胞成瘤性结果

将异源肿瘤细胞分别移植到Rag1-/-小鼠、Rag2-/-小鼠、NOD-SCID小鼠、BALB/c裸小鼠和野生型C57BL/6J小鼠上,观察和比较肿瘤细胞移植后的生长。

结果显示,在Rag1-/-Rag2-/-背景小鼠上,移植肿瘤生长速度最快,且两者之间无显著性差异;在NOD-SCID背景小鼠上的肿瘤生长速度次之;然后是在BALB/c裸小鼠背景;而在C57BL/6J小鼠上则无肿瘤生成(图2A)。Rag1-/-Rag2-/-小鼠上的肿瘤,自肿瘤移植22 d后,显著大于BALB/c裸小鼠和NOD-SCID小鼠上的肿瘤体积(图2A)。

肿瘤组织切片的HE染色结果显示,来自Rag1-/-Rag2-/-小鼠上的移植肿瘤组织和其他品系小鼠上的移植肿瘤组织切片没有显著差异,各组小鼠肿瘤细胞均生长旺盛,肿瘤细胞大小不一,胞核形态比较类似,可见瘤组织均呈巢状或片状生长,细胞间微血管较丰富(图2B)。

Rag1-/-Rag2-/-小鼠相较于传统的异源肿瘤移植受体小鼠品系BALB/cNu裸小鼠和NOD-SCID小鼠,肿瘤生长速度更快。

表1. Rag1-KO(Rag1-EGFP)小鼠的血常规检测。

image.png

表2. Rag1-KO(Rag1-EGFP)小鼠的血生化检测。

image.png

image.png

Fig.8 Detection of mouse IgG(A) and IgM(B) expression in serum by ELISA. 

Abbr. HO, homozygous; WT, wild type; N.D. not detected.



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