A high-quality severe combined immunodeficiency (SCID) rat bioresource.
A high-quality severe combined immunodeficiency (SCID) rat bioresource.
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DOI：
10.1371/journal.pone.0272950
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发表时间：
2022
期刊：
影响因子：
3.7
通讯作者：
中科院分区：
文献类型：
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作者：
免疫缺陷动物是移植外源性组织的有价值的模型;它们被广泛用于许多领域，包括人源化动物模型的创建，以及再生医学和肿瘤学。与小鼠相比，实验室大鼠的体型更大，更容易接受各种组织和器官的移植。考虑到缺乏免疫缺陷大鼠的优质资源，我们使用CRISPR/Cas9基因组编辑系统敲除F344/Jcl大鼠的白细胞介素-2受体γ链基因（Il 2 rg）-单独或与重组激活基因2（Rag 2）一起敲除-以创建研究人员可以自由使用的优质生物资源：用途：严重联合免疫缺陷（SCID）大鼠。我们选择了一只在Il 2 rg（5-bp del）和Rag 2（[1-bp del+2-bp ins]/[7-bp del+2-bp ins]）上都有移码突变的建立者大鼠，然后进行交配以建立免疫缺陷大鼠系。免疫缺陷表型通过Il 2 rg-单敲除（sKO）和Il 2 rg/Rag 2-双敲除（dKO）大鼠中严重胸腺发育不全的存在而初步证实。外周血中血细胞计数的评估显示，sKO和dKO大鼠的白色血细胞计数显著降低，而红细胞计数未受影响。白色血细胞计数减少主要是由淋巴细胞减少引起的。此外，通过流式细胞术分析淋巴细胞群显示，在两种基因敲除大鼠中，B细胞（CD 3-CD 45+）和自然杀伤细胞（CD 3-CD 161+）的数量显著减少。相比之下，T细胞显著减少，但在sKO和dKO大鼠之间显示出略微不同的结果。值得注意的是，我们的免疫缺陷大鼠没有表现出生长迟缓或配子发生缺陷。这种高质量的SCID大鼠资源现在由日本国家生物资源项目管理。我们的SCID大鼠模型已被用于各种研究领域，证明了其作为生物资源的重要性。
Immunodeficient animals are valuable models for the engraftment of exogenous tissues; they are widely used in many fields, including the creation of humanized animal models, as well as regenerative medicine and oncology. Compared with mice, laboratory rats have a larger body size and can more easily undergo transplantation of various tissues and organs. Considering the absence of high-quality resources of immunodeficient rats, we used the CRISPR/Cas9 genome editing system to knock out the interleukin-2 receptor gamma chain gene (Il2rg) in F344/Jcl rats—alone or together with recombination activating gene 2 (Rag2)—to create a high-quality bioresource that researchers can freely use: severe combined immunodeficiency (SCID) rats. We selected one founder rat with frame-shift mutations in both Il2rg (5-bp del) and Rag2 ([1-bp del+2-bp ins]/[7-bp del+2-bp ins]), then conducted mating to establish a line of immunodeficient rats. The immunodeficiency phenotype was preliminarily confirmed by the presence of severe thymic hypoplasia in Il2rg-single knockout (sKO) and Il2rg/Rag2-double knockout (dKO) rats. Assessment of blood cell counts in peripheral blood showed that the white blood cell count was significantly decreased in sKO and dKO rats, while the red blood cell count was unaffected. The decrease in white blood cell count was mainly caused by a decrease in lymphocytes. Furthermore, analyses of lymphocyte populations via flow cytometry showed that the numbers of B cells (CD3- CD45+) and natural killer cells (CD3- CD161+) were markedly reduced in both knockout rats. In contrast, T cells were markedly reduced but showed slightly different results between sKO and dKO rats. Notably, our immunodeficient rats do not exhibit growth retardation or gametogenesis defects. This high-quality SCID rat resource is now managed by the National BioResource Project in Japan. Our SCID rat model has been used in various research fields, demonstrating its importance as a bioresource.
DOI：
10.1292/jvms.18-0107
发表时间：
2018-09-13
期刊：
The Journal of veterinary medical science
影响因子：
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作者：
Tanaka M;Kuramochi M;Nakanishi S;Kuwamura M;Kuramoto T
通讯作者：
Kuramoto T
影响因子：
3.7
作者：
Ito T;Sendai Y;Yamazaki S;Seki-Soma M;Hirose K;Watanabe M;Fukawa K;Nakauchi H
通讯作者：
Nakauchi H