CPC C12N 15/11 (2013.01) [C12N 9/22 (2013.01); C12N 15/902 (2013.01); C12N 2310/20 (2017.05); C12N 2510/02 (2013.01); C12N 2800/80 (2013.01)] | 9 Claims |
1. A genome editing system for site-directed modification of at least one genomic target sequence in the genome of a cell, comprising:
1) an expression construct comprising a coding sequence of a gRNA targeting the at least one genomic target sequence;
2) an expression construct comprising a coding sequence of a CRISPR nuclease; and
3) an expression construct comprising coding sequence for a gRNA targeting a target sequence within the coding sequence of the CRISPR nuclease,
wherein, upon introduction into the cell, said gRNA targeting the at least one genomic target sequence directs the CRISPR nuclease to said at least one genomic target sequence and results in one or more mutations in the genomic target sequence, and the gRNA targeting a target sequence within the coding sequence of the CRISPR nuclease directs the CRISPR nuclease to said target sequence within the coding sequence of the CRISPR nuclease and results in an inactivating mutation of the CRISPR nuclease,
wherein the CRISPR nuclease is a single-base editing CRISPR nuclease comprising the amino acid sequence set forth in SEQ ID NO: 1,
wherein the nucleotide sequence encoding the single-base editing CRISPR nuclease is set forth in SEQ ID NO: 2, and
wherein the target sequence within the coding sequence of the CRISPR nuclease is set forth in SEQ ID NO: 3.
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