A RUNX2-mediated Epigenetic Regulation of the Survival of p53 Defective Cancer Cells

The goal is to identify transcripts that are regulated by RUNX2 in SAOS2 cells. SAOS2 cells were transduced with lentiviruses expressing shLuc, shRUNX2_3 and shRUNX2_4 for 4 days followed by 2ug/ul puromycin selection to remove untransduced cells. A no-virus control was included to monitor the transduction efficiency. These control cells died after 2 days of puromycin selection. The lentivirally transduced cells were more than 80% survived, which indicates a high transduction efficiency. After four more days in puromycin, SAOS2 cells were harvested and subjected to total RNA extraction. 1ug total RNA were used to perform RNA-seq.

本实验的目标为鉴定人成骨肉瘤细胞系SAOS2中受Runt相关转录因子2（RUNX2）调控的转录本。将SAOS2细胞用表达shLuc、shRUNX2_3及shRUNX2_4的慢病毒转导4天，随后以2μg/μl嘌呤霉素进行筛选，以去除未转导的细胞。设置无病毒对照组以监测转导效率，该对照组细胞在嘌呤霉素筛选2天后全部死亡。慢病毒转导的细胞存活率超过80%，表明本次转导效率较高。在含嘌呤霉素的培养基中继续培养4天后，收集SAOS2细胞并进行总RNA提取。取1μg总RNA开展RNA测序（RNA-seq）。