Efficient CRISPR‐based genome editing using tandem guide RNAs and editable surrogate reporters

Cleavage efficiency plays a key role in clustered regularly interspaced short palindromic repeat (CRISPR)‐based gene editing, particularly when the given guide RNA exhibits low cleavage activity. Here, we describe the packaging of tandem guide RNAs and single‐strand annealing‐based surrogate reporter cassettes into the CRISPR/CRISPR‐associated protein 9 vector, which increased gene‐editing efficiency by 4.94–6.31‐fold and simultaneously enriched the proportion of genetically modified cells. This strategy may substantially improve genome‐editing efficiency for demanding applications.