从一名患有导致2型遗传性出血性毛细血管扩张症（HHT）的ACVRL1基因杂合c.1120del18突变的患者中生成2个诱导多能干细胞（iPSC）克隆并进行基因修复。
Generation and genetic repair of 2 iPSC clones from a patient bearing a heterozygous c.1120del18 mutation in the ACVRL1 gene leading to Hereditary Hemorrhagic Telangiectasia (HHT) type 2.
作者信息
Bouma Marga J, Orlova Valeria, van den Hil Francijna E, Mager Hans-Jurgen, Baas Frank, de Knijff Peter, Mummery Christine L, Mikkers Harald, Freund Christian
机构信息
LUMC hiPSC Hotel, Leiden University Medical Center, Leiden, 2333 ZC, The Netherlands; Department of Anatomy and Embryology, Leiden University Medical Center, Leiden, 2333 ZC, The Netherlands.
Department of Anatomy and Embryology, Leiden University Medical Center, Leiden, 2333 ZC, The Netherlands.
出版信息
Stem Cell Res. 2020 Jul;46:101786. doi: 10.1016/j.scr.2020.101786. Epub 2020 May 28.
Fibroblasts from a patient carrying a heterozygous 18bp deletion in exon 8 of the ACVRL1 gene (c.1120del18) were reprogrammed using episomal vectors. The in-frame deletion in ACVRL1 causes the loss of 6 amino acids of the protein, which is associated with Hereditary Hemorrhagic Telangiectasia (HHT) type 2 (Letteboer et al., 2005). CRISPR-Cas9 editing was used to genetically correct the mutation in the induced pluripotent stem cells (iPSCs). The top5-predicted off-target sites were not altered. Patient and isogenic iPSCs showed high pluripotent marker expression, in vitro differentiation capacity into all three germ layers and displayed a normal karyotype. The obtained isogenic pairs will enable proper in vitro disease modelling of HHT (Roman and Hinck, 2017).
利用游离型载体对一名携带ACVRL1基因第8外显子杂合18bp缺失（c.1120del18）的患者的成纤维细胞进行重编程。ACVRL1基因的框内缺失导致该蛋白质6个氨基酸的缺失，这与2型遗传性出血性毛细血管扩张症（HHT）相关（Letteboer等人，2005年）。使用CRISPR-Cas9编辑技术对诱导多能干细胞（iPSC）中的突变进行基因校正。预测的前5个脱靶位点未发生改变。患者和同基因iPSC显示出高多能性标志物表达、体外分化为所有三个胚层的能力，并呈现正常核型。获得的同基因细胞对将有助于对HHT进行合适的体外疾病建模（Roman和Hinck，2017年）。
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