Easy quantitative assessment of genome editing by sequence trace decomposition |
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Authors: | Eva K. Brinkman Tao Chen Mario Amendola Bas van?Steensel |
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Affiliation: | Division of Gene Regulation, Netherlands Cancer Institute, Plesmanlaan 121, 1016 HM Amsterdam, the Netherlands |
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Abstract: | The efficacy and the mutation spectrum of genome editing methods can vary substantially depending on the targeted sequence. A simple, quick assay to accurately characterize and quantify the induced mutations is therefore needed. Here we present TIDE, a method for this purpose that requires only a pair of PCR reactions and two standard capillary sequencing runs. The sequence traces are then analyzed by a specially developed decomposition algorithm that identifies the major induced mutations in the projected editing site and accurately determines their frequency in a cell population. This method is cost-effective and quick, and it provides much more detailed information than current enzyme-based assays. An interactive web tool for automated decomposition of the sequence traces is available. TIDE greatly facilitates the testing and rational design of genome editing strategies. |
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