METHODS AND COMPOSITIONS FOR GENOME-WIDE ANALYSIS AND USE OF GENOME CUTTING AND REPAIR

摘要

Embodiments of the present disclosure relate to compositions and methods for creating and using engineered nucleic acid guided nucleases having a modified gRNA spacer region to modulate targeted gene editing. In certain embodiments, engineered nucleic acid guided nucleases can include a mutated or modified gRNA spacer region to increase editing efficiency and reduce off-targeting of the engineered nuclease. In some embodiments, gRNA spacer regions can be mutated by one or more of a nucleic acid insertion, deletion, point mutation, substitution or other modification to the spacer region to alter recognition and/or binding of a target DNA. In certain embodiments, target DNA can include genomic DNA. In some embodiments, modified spacers of gRNAs can be used in high-throughput editing systems to create specific gRNAs for improved editing efficiency and targeted control over gene editing of use in experimental and therapeutic situations.