CRISPR/Cas9-mediated β-globin gene knockout in rabbits recapitulates human β-thalassemia.
CRISPR/Cas9-mediated β-globin gene knockout in rabbits recapitulates human β-thalassemia.
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CRISPR/Cas9 介导的兔 β 珠蛋白基因敲除再现了人类 β 地中海贫血
DOI：
10.1016/j.jbc.2021.100464
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发表时间：
2021-01
期刊：
影响因子：
--
通讯作者：
Sun X
中科院分区：
文献类型：
--
作者：
Yang Y;Kang X;Hu S;Chen B;Xie Y;Song B;Zhang Q;Wu H;Ou Z;Xian Y;Fan Y;Li X;Lai L;Sun X
β-地中海贫血是一种常染色体隐性遗传性血液疾病，会减少血红蛋白的产生，主要是由 HBB 基因的点突变导致血红蛋白四聚体的 β-珠蛋白链减少或缺失引起的。概括人类疾病表型和基因型的动物模型对于探索病理生理学和新型治疗方法的体内评估很有价值。兔子性情温顺、生命周期短、成本低，使其成为颇具吸引力的动物模型。然而，目前尚无β-地中海贫血兔模型。在这里，我们利用CRISPR/Cas9介导的基因组编辑，对兔β-珠蛋白基因HBB2进行高效点突变，并产生了β-地中海贫血兔模型。血液学和组织学分析表明，基因型嵌合体F0表现出轻度贫血表型，杂合子F1表现出典型的β地中海贫血特征。全血转录组分析显示，与 WT 兔子相比，HBB2 靶向的基因表达发生了改变。 HBB2靶向兔子中高表达的基因在脂质和铁代谢、先天免疫和造血过程中富集。总之，利用 CRISPR 介导的 HBB2 敲除，我们创建了一种β-地中海贫血兔模型，可以准确地重现人类疾病表型。我们相信，该工具对于推进β-地中海贫血及相关并发症的发病机制和新治疗靶点的研究具有重要价值。
β-thalassemia, an autosomal recessive blood disorder that reduces the production of hemoglobin, is majorly caused by the point mutation of the HBB gene resulting in reduced or absent β-globin chains of the hemoglobin tetramer. Animal models recapitulating both the phenotype and genotype of human disease are valuable in the exploration of pathophysiology and for in vivo evaluation of novel therapeutic treatments. The docile temperament, short vital cycles, and low cost of rabbits make them an attractive animal model. However, β-thalassemia rabbit models are currently unavailable. Here, using CRISPR/Cas9-mediated genome editing, we point mutated the rabbit β-globin gene HBB2 with high efficiency and generated a β-thalassemia rabbit model. Hematological and histological analyses demonstrated that the genotypic mosaic F0 displayed a mild phenotype of anemia, and the heterozygous F1 exhibited typical characteristics of β-thalassemia. Whole-blood transcriptome analysis revealed that the gene expression was altered in HBB2-targeted when compared with WT rabbits. And the highly expressed genes in HBB2-targeted rabbits were enriched in lipid and iron metabolism, innate immunity, and hematopoietic processes. In conclusion, using CRISPR-mediated HBB2 knockout, we have created a β-thalassemia rabbit model that accurately recapitulates the human disease phenotype. We believe this tool will be valuable in advancing the investigation of pathogenesis and novel therapeutic targets of β-thalassemia and associated complications.
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影响因子：
20.3
作者：
Magor, Graham W.;Tallack, Michael R.;Perkins, Andrew C.
通讯作者：
Perkins, Andrew C.
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影响因子：
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作者：
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影响因子：
2.3
作者：
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通讯作者：
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影响因子：
64.5
作者：
SKOW, LC;BURKHART, BA;LEWIS, SE
通讯作者：
LEWIS, SE
影响因子：
2.2
作者：
Araki, M;Fan, JL;Watanabe, T
通讯作者：
Watanabe, T