CRISPR-CAS COMPOSITIONS AND METHODS

摘要

Class 2 CRISPR-Cas nucleoprotein complexes are disclosed comprising a Class 2 CRISPR-Cas protein, a CRISPR-Cas associated polynucleotide lacking a spacer element (casPN), and a distinct spacer element sequence polynucleotide (sesPN) comprising a target nucleic acid binding sequence. These complexes are capable of site-directed binding to a target nucleic acid complementary to the target nucleic acid binding sequence of the sesPN. The Class 2 CRISPR-Cas nucleoprotein complexes facilitate site-specific modifications, including cleavage and mutagenesis, of a target nucleic acid sequence. Polynucleotide sequences, expression cassettes, vectors, compositions, and kits for carrying out a variety of methods are also described. Furthermore, the present specification provides guidance for methods of regulating expression of a target nucleic acid sequence, production of genetically modified cells, compositions of modified cells, transgenic organisms, pharmaceutical compositions, as well as a variety of other compositions and methods involving the Class 2 CRISPR-Cas nucleoprotein complexes comprising casPNs, sesPNs, and Cas proteins.