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ResourceVolume 19, Issue 8p1217-1232Open access 1Centre for Stem Cell Biology, School of Biosciences, The University of Sheffield, Sheffield, UK 2Neuroscience Institute, The University of Sheffield, Sheffield, UK 3INSIGNEO Institute, The University of Sheffield, Sheffield, UK 4WiCell Research Institute, Madison, WI, USA 5Department of Human Genetics, University of Chicago, Chicago, IL, USA 6Department of Human Molecular Genetics and Biochemistry, Faculty of Medicine, Tel Aviv University, Tel...
Abstract The CRISPR-Cas system holds great promise in the treatment of diseases caused by genetic variations. As wildtype SpyCas9 is known to generate many off-target effects, its use in the clinic remains controversial due to safety concerns. Several high-fidelity Cas9 variants with greater specificity have been developed using rational design and directed evolution. However, the enhancement of specificity by these methods is limited by factors like selection pressure and library diversity.
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