Figure 4
From: Host Double Strand Break Repair Generates HIV-1 Strains Resistant to CRISPR/Cas9
CRISPR resistant strains have mutations within the gRNA targeted site.
The regions surrounding the CRISPR targeted sites were sequenced 5 weeks after infection. A window of 43 bp is shown and includes 20 bp of gRNA homology (light gray box), 3 bp PAM (dark gray box) and 10 bp of flanking sequence on each side. Red dotted lines indicate the point of cleavage by Cas9. Deletions, total insertions and base substitutions are shown. The total number of mutations in the 43 bp window was set to 100% and the relative contribution of each mutation at each base is indicated as % mutant reads. The frequencies of mutations at flanking sites for all gRNAs are <3%. (a–f) NL4-3 derived CRISPR resistant strains. gRNA-TATA1 n = 1,473,434. gRNA-TATA2 n = 402,382. gRNA-TAR1 n = 655,747. gRNA-TAR2 n = 356,623. gRNA-TAR3 n = 62,704. gRNA-RRE/env n = 908,617. (g–l) R7 derived CRISPR resistant strains. gRNA-TATA1 n = 25,379. gRNA-TATA2 n = 149,465. gRNA-TAR1 n = 207,067. gRNA-TAR2 n = 279,998. gRNA-TAR3 n = 30,232. gRNA-RRE/env n = 749,407.
