COMPOSITIONS AND METHODS OF NUCLEIC ACID-TARGETING NUCLEIC ACIDS
This disclosure provides for compositions and methods for the use of nucleic acid-targeting nucleic acids and complexes thereof.
1 - 26 . (canceled)
27 . A method for delivering a donor polynucleotide to a double-stranded break at a target site, comprising:
contacting a DNA sequence comprising the target site with an engineered nucleic acid targeting nucleic acid comprising a crRNA and tracrRNA wherein (i) the engineered nucleic acid targeting nucleic acid is associated in a complex with a Cas9 protein to form a crRNA-tracrRNA-Cas9 complex, (ii) the tracrRNA comprises a 3′ end, (iii) the tracrRNA further comprises a hybridizing sequence at the 3′ end, (iv) the hybridizing sequence comprises a nucleic acid sequence that hybridizes to the donor polynucleotide, and (v) the crRNA-tracrRNA-Cas9 complex binds to the DNA sequence comprising the target site and cuts the target site to generate a double-strand break and the binding of the crRNA-tracrRNA-Cas9 complex delivers the donor polynucleotide to the double-stranded break at the target site.