Cell Death Caused by Single-Stranded Oligodeoxynucleotide-Mediated Targeted Genomic Sequence Modification
Cell Death Caused by Single-Stranded Oligodeoxynucleotide-Mediated Targeted Genomic Sequence Modification
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DOI：
10.1089/oli.2009.0191
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发表时间：
2009-09-01
期刊：
影响因子：
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通讯作者：
Huang, Jian-Dong
中科院分区：
文献类型：
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作者：
Liu, Chenli;Wang, Zai;Huang, Jian-Dong
由单链寡脱氧核苷酸（ssODN）指导的靶向基因修复为生物技术和基因治疗提供了一种有前途的工具。然而，该方法目前受到其低频率的修复事件、可变性和“校正的”细胞的低活力的限制。在这项研究中，我们表明，在ssODN介导的基因修复反应，一个显着的人口校正细胞未能分裂，更容易发生凋亡，标记的加工半胱天冬酶和PARP-1。此外，我们发现ssODN介导的基因修复引发的凋亡性细胞死亡在很大程度上不依赖于ATM/ATR激酶。此外，我们还研究了错配修复（MMR）蛋白在这种“校正反应诱导的”细胞死亡中的潜在参与。结果显示，虽然缺陷MMR大大提高了基因校正的效率，但损害MMR系统并不产生任何可行的校正克隆，这表明MMR机制虽然在确定ssODN-定向修复中起关键作用，但并不参与观察到的细胞遗传毒性反应。
Targeted gene repair directed by single-stranded oligodeoxynucleotides (ssODNs) offers a promising tool for biotechnology and gene therapy. However, the methodology is currently limited by its low frequency of repair events, variability, and low viability of "corrected" cells. In this study, we showed that during ssODN-mediated gene repair reaction, a significant population of corrected cells failed to divide, and were much more prone to undergo apoptosis, as marked by processing of caspases and PARP-1. In addition, we found that apoptotic cell death triggered by ssODN-mediated gene repair was largely independent of the ATM/ATR kinase. Furthermore, we examined the potential involvement of the mismatch repair (MMR) proteins in this "correction reaction-induced" cell death. Result showed that while defective MMR greatly enhanced the efficiency of gene correction, compromising the MMR system did not yield any viable corrected clone, indicating that the MMR machinery, although plays a critical role in determining ssODN-directed repair, was not involved in the observed cellular genotoxic responses.