Article
Amplification-free long-read sequencing reveals unforeseen CRISPR-Cas9 off-target activity.
Genome biology - 1 Dec 2020
Höijer Ida, Johansson Josefin, Gudmundsson Sanna, Chin Chen-Shan, Bunikis Ignas, Häggqvist Susana, Emmanouilidou Anastasia, Wilbe Maria, den Hoed Marcel, Bondeson Marie-Louise, Feuk Lars, Gyllensten Ulf, Ameur Adam
Abstract excerpt
BACKGROUND: One ongoing concern about CRISPR-Cas9 genome editing is that unspecific guide RNA (gRNA) binding may induce off-target mutations. However, accurate prediction of CRISPR-Cas9 off-target activity is challenging. Here, we present SMRT-OTS and Nano-OTS, two novel, amplification-free, long-read sequencing protocols for detection of gRNA-driven digestion of genomic DNA by Cas9 in vitro. RESULTS: The methods...
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