骨架修饰:pINDUCER11 (miR-RUG) was made by first digesting pINDUCER10 with XbaI and PacI to remove the rtTA through the puromycin resistance gene. The rtTAIRES segment was removed from pIndmir3Luc-miR by XbaI and NotI digest. eGFP was PCR amplified from MSCV-N-eGFP-ENTR using primers 5′-TAATACATGT ATCGATCCACCATGGTGAGCAAGGGC and 5′-GATCTTAATTAA TTACTTGTACAGCTCGTCCATGCCG. The PCR product was digested with PciI and PacI. The vector, rtTA-IRES, and eGFP were then ligated and sequence verified.