Meganuclease-mediated virus self-cleavage facilitates tumor-specific virus replication.
Meganuclease-mediated virus self-cleavage facilitates tumor-specific virus replication.
复制标题
大范围核酸酶介导的病毒自裂解促进肿瘤特异性病毒复制
DOI：
10.1038/mt.2013.117
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发表时间：
2013
期刊：
影响因子：
--
通讯作者：
Kühnel F.
中科院分区：
文献类型：
--
作者：
Gürlevik E;Schache P;Goez A;Kloos A;Woller N;Armbrecht N;Manns M;Kubicka S;Kühnel F.
巨核酶可以特异性地切割长DNA序列基序，这一特性使它们成为活细胞基因工程的理想工具。在一项概念验证研究中，我们研究了使用巨核酶I- sce I进行靶向病毒自我破坏以产生高特异性溶瘤病毒。为此，我们为溶瘤腺病毒提供了一种分子电路，该分子电路通过表达I- sce I选择性地响应p53的激活，随后通过病毒基因组内的异源I- sce I识别位点导致病毒DNA的自我破坏。我们观察到病毒复制和细胞裂解在p53-正常细胞中被有效地破坏，而在p53-功能失调的肿瘤细胞中没有。通过检测特定的切割产物，证实了I- sce I活性导致病毒DNA在细胞内的有效加工。病毒破坏不会干扰E1A水平，这表明功能性病毒基因组的减少是条件复制的主要原因。因此，当通过靶向转录抑制进一步抑制E1A表达时，肿瘤特异性复制进一步增强。最后，我们在体外和体内证明了表达I-Sce i的病毒对p53依赖性肿瘤的溶解作用，并证明了对肿瘤生长的有效抑制。总之，巨核酶介导的病毒切割是一种很有前途的方法，可以为溶瘤病毒提供具有吸引力的安全性。
Meganucleases can specifically cleave long DNA sequence motifs, a feature that makes them an ideal tool for gene engineering in living cells. In a proof-of-concept study, we investigated the use of the meganuclease I-Sce I for targeted virus self-disruption to generate high-specific oncolytic viruses. For this purpose, we provided oncolytic adenoviruses with a molecular circuit that selectively responds to p53 activation by expression of I-Sce I subsequently leading to self-disruption of the viral DNA via heterologous I-Sce I recognition sites within the virus genome. We observed that virus replication and cell lysis was effectively impaired in p53-normal cells, but not in p53-dysfunctional tumor cells. I-Sce I activity led to effective intracellular processing of viral DNA as confirmed by detection of specific cleavage products. Virus disruption did not interfere with E1A levels indicating that reduction of functional virus genomes was the predominant cause for conditional replication. Consequently, tumor-specific replication was further enhanced when E1A expression was additionally inhibited by targeted transcriptional repression. Finally, we demonstrated p53-dependent oncolysis by I-Sce I-expressing virusesin vitroandin vivo, and demonstrated effective inhibition of tumor growth. In summary, meganuclease-mediated virus cleavage represents a promising approach to provide oncolytic viruses with attractive safety profiles.
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影响因子：
4.2
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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病毒和非病毒因素导致肿瘤和组织特异性启动子依赖性溶瘤腺病毒的非特异性复制。
影响因子：
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