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Technology

The technology is a miniature CRISPR nuclease that is target specific, compact in structure, and made up of a small number of amino acids (<800-1000). The nuclease targets DNA and is directed by a guide RNA (gRNA) to a target nucleic acid sequence, which is commonly a protospacer adjacent motif (PAM). The gRNA can be a single-guide RNA, which is a blend of two non-coding RNAs: a synthetic CRISPR RNA (crRNA) and a trans-activating CRISPR RNA (tracrRNA). This system allows for the insertion or deletion of base pairs in DNA. The target specific nuclease cleaves the DNA at a target site, leaving overhangs on both ends of the DNA. Then, in nucleotide insertion, a nucleotide is added complementary to the overhanging nucleotide on both ends. In deletion, the overhanging nucleotides on both ends are removed. Finally, the DNA ends are ligated together to complete the insertion or deletion of base pairs.