CRISPR ENZYMES, METHODS, SYSTEMS AND USES THEREOF

The present invention provides novel systems, methods and compositions for making and using recombinantly engineered novel Cas9 enzymes optimized for human cells, for nucleic acid targeting and manipulation. The present invention is based on the discovery of novel Cas9 enzymes from Streptococcus equinus ATCC 33317, Enterococcus hirae strain F1129E, Streptococcus equinus strain AG46, Staphylococcus simulans strain 19, Streptococcus intermedius B196 strain G1552, Streptococcus sanguinis SK330, Streptococcus sp. C150, Streptococcus oralis subsp. oralis strain RH_1735_08, Streptococcus oralis SK313, Staphylococcus warneri strain 691, Staphylococcus stiuri strain SNUC 2430, Streptococcus gallolyticus strain AM24-4, Lactobacillus kullabergensis strain Biut2, and Streptococcus suis strain LSS83 bacteria that were codon-optimized and recombinantly produced for use in human cells. In some embodiments, novel Cas9 enzymes can be used for base editing. In some embodiments, the novel engineered Cas9 enzymes are used to treat human diseases.