| Screening of Efficient siRNA Target Sites Directed against Gatekeeper Genes for DNA Repair |
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| 作者姓名： | 任精华 林菊生 董旭 徐冬 陈琼 刘瑶 常莹 姚津剑 韩思源 |
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| 作者单位： | Institute of Liver Diseases Tongji Hospital Tongji Medical College Huazhong University of Science and Technology，Institute of Liver Diseases Tongji Hospital Tongji Medical College Huazhong University of Science and Technology，Institute of Liver Diseases Tongji Hospital Tongji Medical College Huazhong University of Science and Technology，Institute of Liver Diseases Tongji Hospital Tongji Medical College Huazhong University of Science and Technology，Institute of Liver Diseases Tongji Hospital Tongji Medical College Huazhong University of Science and Technology，Institute of Liver Diseases Tongji Hospital Tongji Medical College Huazhong University of Science and Technology，Institute of Liver Diseases Tongji Hospital Tongji Medical College Huazhong University of Science and Technology，Institute of Liver Diseases Tongji Hospital Tongji Medical College Huazhong University of Science and Technology，Institute of Liver Diseases Tongji Hospital Tongji Medical College Huazhong University of Science and Technology，Wuhan 430030 China，Wuhan 430030 China，Wuhan 430030 China，Wuhan 430030 China，Wuhan 430030 China，Wuhan 430030 China，Wuhan 430030 China，Wuhan 430030 China，Wuhan 430030 China |
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| 摘 要： | DNA double-strand breaks (DSBs) are common le-sions that occur in cells. They are caused by exogenous sources such as ionizing radiation, and by endogenous sources such as radicals generated during metabolic processes1]. In mammalian cells, DSBs are repaired ei-ther by the homologous recombination (HR) pathways or by the non-homologous end joining (NHEJ)2] pathways to maintain the fidelity of human genome. The basic mechanism and factor requirements of the two pathways are different. V…
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| 关 键 词： | 基因表达 干扰技术 生物 研究 |
| 收稿时间： | 2006-11-18 |
| 修稿时间： | 2006-11-18 |
Screening of efficient siRNA target sites directed against gatekeeper genes for DNA repair |
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| Jinghua Ren M.D., Ph.D.,Jusheng Lin,Xuyang Dong,Dong Xu,Qiong Chen,Yao Liu,Ying Chang,Jinjian Yao,Siyuan Han.Screening of Efficient siRNA Target Sites Directed against Gatekeeper Genes for DNA Repair[J].Journal of Zuazhong University of Science and Technology: Medical Edition,2006,26(6):640-643. |
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| Authors: | Jinghua Ren MD PhD Jusheng Lin Xuyang Dong Dong Xu Qiong Chen Yao Liu Ying Chang Jinjian Yao Siyuan Han |
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| Affiliation: | Institute of Liver Diseases, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology,Wuhan 430030, China |
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| Abstract: | To investigate the RNA interference (RNAi) effect induced by vector-derived small in-terfering RNA (siRNA) targeting the three gatekeeper genes (Rad52, Ku70, Ku80) and screen the more effective target sites from candidates for further research, by using siRNA design tools online, we selected 2 candidate sequences directed to every gatekeeper gene. According to the sequences, six vector-derived siRNAs (denoted psiRNA1-6) and one mocking psiRNA7 were constructed. Among them, psiRNA1 and psiRNA2 targeted Rad52, psiRNA3 and psiRNA4 to Ku70, psiRNA5 and psiRNA6 to Ku80. The mocking psiRNA7 was used as control. After sequence identification, the seven plasmids were transfected into HepG2 cell line. siRNA-induced silencing of gatekeeper genes was determined by using RT-PCR at RNA level and Western Blot at protein level. The results showed that the six plasmids specifically targeting the coding region of gatekeeper genes were successfully designed and constructed. To some extent, the six plasmids could reduce the expression of target gene. Comparatively, the plasmid-derived siRNA psiRNA1, psiRNA4 and psiRNA5 were more effective than their counterparts. The results suggest that the gene silencing efficiency of siRNA is different, depending on their targeted region, and siRNA may provide us with practical tools for further study on the three gatekeeper genes, i.e. Rad52, Ku70, Ku80. |
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| Keywords: | gatekeeper gene RNA interference vector-derived siRNA |
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