Zhou J.-Y.,Guangzhou First Municiple People Hospital |
Deng H.,Guangzhou First Municiple People Hospital |
Wang G.-L.,Guangzhou First Municiple People Hospital |
Zhang Y.-P.,Guangzhou First Municiple People Hospital |
And 3 more authors.
Journal of Leukemia and Lymphoma | Year: 2012
Objective To establish stable HL-60 cell line with stable RYBP gene silencing using lentivirus-mediated RNA interference. Methods Five special shRNAs for RYBP gene were cloned to lentivirus vector. Recombinate lentivirus vectors were packed into lentivirus, which were used to infect HL-60 cells, and took empty vector and non-specific shRNA as control groups. Stable infected cells were selected with puromycin in 8 μg/ml concentration. The expression levels of RYBP were analyzed by Western blot, and confirmed the most effective RYBP shRNA. Then the level of mRNA was analyzed by real-time PCR. Results Stable infected cells were selected by puromycin successfully. Comparing to control groups, the expression of RYBP were reduced at different degrees (P < 0.01). And RYBP shRNA2 took the most silencing effect, the RYBP mRNA was decreased by more than 95 % (P < 0.05). Conclusion The shRNA2 targeting RYBP gene can effectively inhibit the expression of RYBP. HL-60 cell line with stable RYBP gene silencing were constructed successfully, which had provided experiment fundament for further studying the function of RYBP.