同源重组细胞基因敲除与siRNA基因沉默的效果比较  

Comparison of Effect between Homologous Recombinant Gene Knockout and siRNA Gene Silence in Cell Lines

作　　者：李庆华 金薇娜 张华梅 茹永新 庞天翔 

机构地区：[1]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

出　　处：《中国实验血液学杂志》2010年第1期122-126,共5页Journal of Experimental Hematology

基　　金：天津市自然科学基金资助项目(编号08JCZDJC19100,09JCZDJC17300)

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。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 growth retardation in Sam68 gene knocked-out cell line was observed due to elongation of the G2/M phase, but which could not be found in Sam68 gene silencing cell line. It is concluded that in accordance with study of protein function in living cells, use of gene knockout technique for cell line can provide the experimental results more real than those resulting from gene silence technique.

关 键 词：基因敲除 基因沉默 Sam68 DT40 

分 类 号：Q789[生物学—分子生物学]