siDNMT1 increases γ-globin expression in chemical inducer of dimerization (CID)-dependent mouse βYAC bone marrow cells and in baboon erythroid progenitor cell cultures.
siDNMT1 increases γ-globin expression in chemical inducer of dimerization (CID)-dependent mouse βYAC bone marrow cells and in baboon erythroid progenitor cell cultures.
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DOI：
10.1016/j.exphem.2010.10.003
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发表时间：
2011-01
影响因子：
2.6
通讯作者：
Lavelle, Donald
中科院分区：
文献类型：
--
作者：
Banzon, Virryan;Ibanez, Vinzon;Vaitkus, Kestis;Ruiz, Maria Armila;Peterson, Kenneth;DeSimmone, Joseph;Lavelle, Donald
进行这些研究以检验DNMT 1是维持成人期红系细胞中DNA甲基化和抑制γ-珠蛋白基因所必需的假设。在酵母人工染色体（βYAC）背景下含有人β-珠蛋白基因位点的化学二聚化诱导剂（CID）依赖性多能小鼠骨髓（BM）细胞和来源于CD 34+狒狒BM细胞的红系祖细胞原代培养物中，通过靶向DNMT 1的siRNA核转染降低DNMT 1水平。通过真实的时间PCR测量降低的DNMT 1水平对珠蛋白基因表达的影响，并且通过珠蛋白链的生物合成放射性标记随后通过HPLC分析来确定对原代红系祖细胞培养物中的珠蛋白链合成的影响。通过亚硫酸氢盐序列分析确定对DNA甲基化的影响。用siDNMT 1处理的细胞中DNMT 1水平的降低与γ-珠蛋白mRNA表达的增加、培养的红系祖细胞中γ/γ+β链比率的增加以及γ-珠蛋白启动子的DNA甲基化的降低相关。在用DNA甲基转移酶抑制剂的药理学抑制剂地西他滨处理的细胞中观察到类似的效果。DNMT 1是维持γ-珠蛋白基因启动子DNA甲基化和抑制成熟期红系细胞中γ-珠蛋白基因表达所必需的。
These studies were performed to test the hypothesis that DNMT1 is required for maintenance of DNA methylation and repression of the γ-globin gene in adult stage erythroid cells. DNMT1 levels were reduced by nucleofection of siRNA targeting DNMT1 in chemical-inducer-of-dimerization (CID)-dependent multipotential mouse bone marrow (BM) cells containing the human β-globin gene locus in the context of a yeast artificial chromosome (βYAC) and in primary cultures of erythroid progenitor cells derived from CD34+ baboon BM cells. The effect of reduced DNMT1 levels on globin gene expression was measured by real time PCR and the effect on globin chain synthesis in primary erythroid progenitor cell cultures was determined by biosynthetic radiolabelling of globin chains followed by HPLC analysis. The effect on DNA methylation was determined by bisulfite sequence analysis. Reduced DNMT1 levels in cells treated with siDNMT1 were associated with increased expression of γ-globin mRNA, an increased γ/γ+β chain ratio in cultured erythroid progenitors, and decreased DNA methylation of the γ-globin promoter. Similar effects were observed in cells treated with decitabine, a pharmacological inhibitor of DNA methyltransferase inhibitor. DNMT1 is required to maintain DNA methylation of the γ-globin gene promoters and repress γ-globin gene expression in adult-stage erythroid cells.
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影响因子：
3.6
作者：
Lavelle, DE
通讯作者：
Lavelle, DE
影响因子：
20.3
作者：
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Dnmt1 和 Dnmt3a 维持 DNA 甲基化并调节成人前脑神经元的突触功能
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作者：
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通讯作者：
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影响因子：
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作者：
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20.3
作者：
Fathallah, Hassana;Weinberg, Rona S.;Atweh, George F.
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