Reactivation of silenced, virally transduced genes by inhibitors of histone deacetylase
Reactivation of silenced, virally transduced genes by inhibitors of histone deacetylase
复制标题
DOI：
10.1073/pnas.94.11.5798
复制
发表时间：
1997-05-27
影响因子：
11.1
通讯作者：
Townes, TM
中科院分区：
文献类型：
--
作者：
Chen, WY;Bailey, EC;Townes, TM
逆转录病毒和腺相关病毒序列可以显著沉默小鼠中的转基因表达。我们现在报道，当细胞在没有选择的情况下生长时，这种抑制也发生在稳定感染的HeLa细胞中。在培养60天后，转导的lacZ基因（rAAV/CMVlacZ）的表达在超过90%的细胞中沉默。令人惊讶的是，高水平表达可以通过用丁酸钠或胱抑素A处理细胞而不是用5-氮杂胞苷处理细胞来重新激活。当分离具有rAAV/CMVlacZ整合拷贝的细胞克隆时，80%的克隆中lacZ表达沉默;然而，通过丁酸盐或曲马多菌素A处理，所有沉默的克隆中lacZ表达被重新激活。这两种药物也重新激活了一个沉默的珠蛋白基因结构（rAAV/HS 2 α β（AS 3））中，曲古抑菌素A是组蛋白脱乙酰酶的特异性抑制剂;因此，我们认为药物治疗后组蛋白的超乙酰化改变了整合的病毒序列上的染色质结构，并解除了转导基因的抑制。沉默的，有效的病毒基因转移和随后的药物治疗以解除抑制可以为治疗各种遗传和感染性疾病提供强有力的组合。
Retroviral and adeno-associated viral sequences can dramatically silence transgene expression in mice. We now report that this repression also occurs in stably infected HeLa cells when the cells are grown without selection, Expression of a transduced lacZ gene (rAAV/CMVlacZ) is silenced in greater than 90% of cells after 60 days in culture, Surprisingly, high-level expression can be reactivated by treating the cells with sodium butyrate or trichostatin A but not with 5-azacytidine. When cell clones with integrated copies of rAAV/CMVlacZ were isolated, lacZ expression was silenced in 80% of the clones; however, lacZ expression was reactivated in all of the silenced clones by treatment with butyrate or trichostatin A. The two drugs also reactivated a silenced globin gene construct (rAAV/HS2 alpha beta(AS3)) in stably infected K562 cells, Trichostatin A is a specific inhibitor of histone deacetylase; therefore, we propose that hyperacetylation of histones after drug treatment changes the structure of chromatin on integrated viral sequences and relieves repression of transduced genes, The reactivation of silenced, transduced genes has implications for gene therapy, Efficient viral gene transfer followed by drug treatment to relieve suppression may provide a powerful combination for treatment of various genetic and infectious diseases.