[Comparison of effect between homologous recombinant gene knockout and siRNA gene silence in cell lines].
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.
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.
Qing-Hua
QH
Li
Wei-Na
WN
Jin
Hua-Mei
HM
Zhang
Yong-Xin
YX
Ru
Tian-Xiang
TX
Pang
D000818:Animals; D045744:Cell Line, Tumor; D002645:Chickens; D015972:Gene Expression Regulation, Neoplastic; D055786:Gene Knockout Techniques; D020868:Gene Silencing; D018390:Gene Targeting; D010957:Plasmids; D034741:RNA, Small Interfering / Q000235:genetics*; D014162:Transfection
D034741:RNA, Small Interfering