Figure 3
From: Targeted editing of the PSIP1 gene encoding LEDGF/p75 protects cells against HIV infection
HDR template design and screening. (a) Schematic representation of exon12 sequence of PSIP1 gene harboring D366 residue. Below the designed HDR template as a ssDNA oligo is shown with red characters indicating the sites of nucleotide substitutions, generating a VspI restriction site at D366N and deleting the PAM site. (b) Sanger sequencing results for the HEK293T_LEDGF D366N clone that was identified, showing consensus between HDR template and allele 1 (substitution indicated in red). The sequence for allele 2 shows a 8-nucleotide deletion (“−” indicates deletion) and the translation (capital letters below) resulting in a premature stop codon (underlined). (c) Western blot analysis showing the LEDGF/p75 levels in D366N clone compared to WT levels.
