[Effect of Sry silencing by siRNA on the expression of sex determining genes in mouse embryos].
In order to investigate Sry regulation network in the development of male embryo, we inhibited the Sry gene expression by RNAi and then examined expression of other sex-related genes. Six genes (Sox9, Wt1, Sf1, Dax1, Gata4 and Amh), which are suggested to be closely related to Sry regulation were studied. Two siRNA expression vectors pSilencer4.1/Sry217 and pSilencer4.1/Sry565 were constructed and injected into gestated mouse through tail vein at 9.5 day of conception (dpc). The inhibition efficiency of Sry and the expression of other six genes were examined in male embryos at 11.5 dpc by RT-PCR and Western-blot. Expressions of the other six genes were analyzed by fluorescence quantity PCR. The results indicated both the pSilencer4.1/Sry217 and pSilencer4.1/Sry565 could inhibit significantly increased after Sry silencing. In contrast, no significant changes were observed in the expression of Sf1, Amh, Gata4, Dax1 and Sox9 when silencing Sry by siRNA. Our results suggested that the Wt1 transcription was regulated by Sry, whereas the Sox9 expression is not directly regulated by Sry in the development of genital ridge.
Wu
Ning
N
Institute of Animal Science, Chinese Academy of Agricultural Sciences, Beijing 100094, China. wuning2283720@126.com
Lin
Xiu-Kun
XK
Liao
Bing
B
DU
Wei-Hua
WH
Han
Feng-Tong
FT
Zhao
Jin-Hong
JH
D000818:Animals; D015153:Blotting, Western; D004622:Embryo, Mammalian; D005260:Female; D055785:Gene Knockdown Techniques; D053263:Gene Regulatory Networks; D008297:Male; D051379:Mice; D016133:Polymerase Chain Reaction; D011247:Pregnancy; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000378:metabolism*; D019849:Sex Determination Processes; D051876:Sex-Determining Region Y Protein / Q000172:deficiency* / Q000235:genetics*; D015533:Transcriptional Activation
D034741:RNA, Small Interfering; D051876:Sex-Determining Region Y Protein