Inhibitory effect of RNA interference of MSTN gene expression on the downstream genes in Schizopygopsis pylzovi / 中国实验动物学报

ABSTRACT

Objective To investigate the silencing effect of RNA interference on MSTN gene ( myostatin, MSTN) expression, and detect the effects on the downstream genes in Schizopygopsis pylzovi. Methods To construct the recombi?nant adenovirus vector 1P3 (DSP MSTN 273+250+1737) and 1P2 (DSP MSTN 195+ 1670) for RNA interference of the MSTN gene in Schizopygopsis pylzovi, and to conduct the RNA interference in vivo experiment by injecting the vector in?to the muscle tissue of Schizopygopsis pylzovi. Real?time PCR and Western blotting were used to evaluate the silencing effects on MSTN gene expression, and to detect the regulatory function of M?CK at gene transcription level after RNA inter?ference of the MSTN gene. Results The result of real?time PCR showed that compared with the HK team ( Virus general negative control group) and N team (blank control group), the 1P3 had significant interference effect on the MSTN gene transcription in Schizopygopsis pylzovi (P<0?05), with an inhibition rate of 53?5%, but the 1P2 had no significant inter?ference effect on the MSTN gene transcription. The result of Western blotting was consistent with the results of real?time PCR. At the same time, after the 1P3 interference, the level of MSTN gene transcription was declined, and the level of M? CK gene expression was significantly increased. Conclusions Our results demonstrate that the expression of MSTN gene can be effectively suppressed, and the expression of M?CK gene can be up?regulated through the RNA interference. There?fore, it proves that MSTN gene can inhibit the transcription of M?CK gene in Schizopygopsis pylzovi, and reveals the regula?tory role of MSTN gene in the muscle growth and development in the plateau fish Schizopygopsis pylzovi.