Figure 5
From: Targeted editing of the PSIP1 gene encoding LEDGF/p75 protects cells against HIV infection
HIV replication is severely affected in SupT1_LEDGF KO cells. (a) Schematic representation of a detailed zoom of exon12 sequence of the PSIP1 gene harboring D366. Sanger sequencing of the indel profile generated in the region targeted by gRNA1 (“−” indicates deletion, substitution indicated in red) and the predicted changes in the amino acid sequence of exon 12 shown on the right. (b) Western analysis of LEDGF/p75 protein in the 3 isolated LEDGF KO cell lines. (c) PSIP1 mRNA expression relative to β-actin in the respective SupT1_LEDGF KO clones as determined by qPCR. Error bars represent SD. Student’s t test was performed using GraphPad Prism 7.0 software. Sample means were considered significantly different from the WT control at p < 0.05 (*). (d) HIV-1 replication assay. The respective cell lines were challenged with the laboratory HIVNL4.3 strain at a final concentration of 5.0*102 pg p24. Viral replication was monitored by daily sampling of p24 in the cell culture supernatant. The graph shows a representative infection experiment out of three independent trials.
