| 同源重组细胞基因敲除与siRNA基因沉默的效果比较 |
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| 引用本文： | 李庆华,金薇娜,张华梅,茹永新,庞天翔. 同源重组细胞基因敲除与siRNA基因沉默的效果比较[J]. 中国实验血液学杂志, 2010, 18(1): 122-126 |
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| 作者姓名： | 李庆华 金薇娜 张华梅 茹永新 庞天翔 |
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| 作者单位： | 中国医学科学院、北京协和医学院血液学研究所、血液病医院,实验血液学国家重点实验室,天津,300020 |
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| 基金项目： | 天津市自然科学基金资助项目(编号08JCZDJC19100,09JCZDJC17300) |
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| 摘 要： | 本研究比较细胞系同源重组基因敲除(gene knockout)与siRNA基因沉默(gene silence)二种技术方法用于研究蛋白功能方面的效果差异。对鸡B淋巴细胞瘤来源的DT40细胞系,应用基因打靶技术分离出Sam68基因缺失的细胞株,并且利用稳定表达siRNA的质粒pSilencer-Sam68得到Sam68基因沉默的细胞株。与野生型细胞株相比,对这些细胞株进行Sam68的功能解析。结果表明:Sam68基因缺失细胞表现出明显的生长速度迟缓,通过细胞周期检测揭示这些细胞生长速度延迟是由于细胞周期中的G2/M期延长;但在Sam68基因沉默细胞中却没有看到这些变化。结论:针对活细胞中的蛋白功能研究,应用细胞系基因敲除技术比siRNA基因沉默技术能够得到更加清晰的实验结果。
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| 关 键 词： | 基因敲除 基因沉默 Sam68 DT40 |
Comparison of Effect between Homologous Recombinant Gene Knockout and siRNA Gene Silence in Cell Lines |
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| LI Qing-Hua,JIN Wei-Na,ZHANG Hua-Mei,RU Yong-Xin,PANG Tian-Xiang. Comparison of Effect between Homologous Recombinant Gene Knockout and siRNA Gene Silence in Cell Lines[J]. Journal of experimental hematology, 2010, 18(1): 122-126 |
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| Authors: | LI Qing-Hua JIN Wei-Na ZHANG Hua-Mei RU Yong-Xin PANG Tian-Xiang |
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| Affiliation: | (State key Laboratory of Experimental Hematology, Institute of Hematology and Hospital of Blood Diseases, Chinese Academy of Medical Sciences and Peking Union Medical College, Tianjin 300020, China) |
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| Abstract: | The objective of this study was to compare the effects between knocking-out Sam68 gene by homologous recombination method and silencing the gene by siRNA silencing technique in DT40 cell line.Gene targeting technique was used to isolate Sam68 gene-deleted chicken DT40 cells.Meanwhile,Sam68 gene silencing cells was obtained by using stable expression of siRNA plasmid pSilencer-Sam68.Then,the function of these two cell lines were analyzed by comparing with wild-type DT40 cell line.The results showed that the ... |
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| Keywords: | gene knockout gene knockdown Sam68 DT40 |
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