Chapter 18, Problem 26P

One problem that researchers sometimes encounter when editing genomes with CRISPR/Cas9 is that one or more loci other than the intended target can be recognized by Cas9/sgRNA and cleaved. Part of the reason is that single base pair mismatches between the target site and the sgRNA in the 5′-most half of the 20 bp DNA/RNA hybrid do not prevent Cas9 cleavage of the target site. How could scientists use bioinformatics to avoid such off-target effects?