Incoming human papillomavirus 16 genome is lost in PML protein-deficient HACAT keratinocytes.. Homo sapiens

Human papillomaviruses (HPVs) target PML nuclear bodies during infectious entry and PML protein is important for efficient transcription of incoming viral genome.We used shRNA to knockdown PML protein in HaCaT keratinocytes to further investigate the role of PML protein in HPV entry. We used microarrays to compare the transcriptome of PML protein-deficient with vector control-transduced HaCaT cells. Overall design: HaCaT cells were transduced with lentiviruses expresing shRNA directed against PML or scrambled control. Stable cell lines were then selected with puromycin. A PML-deficient single-cell clone and vector control cells were used to isolate total RNA.

人类乳头瘤病毒（Human papillomaviruses, HPVs）在感染入侵过程中靶向PML核体（PML nuclear bodies），且PML蛋白对于入侵病毒基因组的高效转录至关重要。为进一步探究PML蛋白在HPV入侵过程中的作用，我们在HaCaT角质形成细胞（HaCaT keratinocytes）中利用短发夹RNA（short hairpin RNA, shRNA）敲低PML蛋白的表达。我们采用基因微阵列（microarrays）对比了PML蛋白缺陷型与载体对照转导的HaCaT细胞的转录组。整体实验设计：将HaCaT细胞用靶向PML的shRNA或乱序对照的慢病毒进行转导，随后通过嘌呤霉素（puromycin）筛选获得稳定细胞株；选取一株PML缺陷型单细胞克隆株与载体对照细胞，提取总RNA。