Figure 7
From: Targeted editing of the PSIP1 gene encoding LEDGF/p75 protects cells against HIV infection
Characterization of SupT1_LEDGF D366N. (a) Sanger sequencing results showing consensus between HDR template and allele 1. The HDR template is shown with red characters indicating the sites of nucleotide substitutions, generating a VspI restriction site at D366N and deleting the PAM site. The sequence for allele 2 shows a 22-nucleotide deletion gRNA1 (“−” indicates deletion), resulting in KO of the PSIP1 allele. (b) Western blot analysis for the SupT1_LEDGF D366N clone using C-terminal specific LEDGF antibody, αLEDGF480–530. SupT1_WT and KO cells are included as controls. (c) Immunocytochemical analysis of LEDGF D366N protein (green) in SupT1_LEDGF D366N cells (bottom panel) recapitulates that of LEDGF/p75 in SupT1_WT cells (top panel). Phalloidin (white) was used to stain F-actin. Scale Bar: 10 µm.
