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Figure 1 | BMC Genomics

Figure 1

From: A gene-rich, transcriptionally active environment and the pre-deposition of repressive marks are predictive of susceptibility to KRAB/KAP1-mediated silencing

Figure 1

Isolation of thousands of KRAB/KAP1 recruitment sites in genes with variable silencing phenotypes. (A) Experimental setup used to target tTRKRAB to endogenous genes through the retroviral vector-based promoter trapping/silencing (TrapSil) system. TetO-containing gene traps, which carry the promoterless GFP-puroR fusion reporter, are only expressed if they trap an actively transcribed gene. In the absence of doxycycline (Dox-), binding of the ectopic tTRKRAB repressor to TetO mediates silencing by recruiting KAP1 and associated heterochromatin-inducing factors. (B) Method used for isolating TrapSil HeLa cell subpopulations based on the effect of tTRKRAB binding on trapped gene expression. Repressible (REP) clones exhibit silencing of the reporter gene upon tTRKRAB binding, whereas reporter transcription of non-repressible clones (NREP) remains unaltered in this condition. (C) This table shows the outcome of the proviral integrant mapping and their distribution relative to genes. Mapping to the genome was performed with FetchGWI. The determination of intragenic integration sites was based on UCSC known genes. LVtotal and MLVtotal encompass all LV- and MLV -integrants. In addition, LV- and MLV- REP and NREP describe the repressible and non-repressible subsets of each vector type.

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