Dual guide CRISPR hybrid DNA/RNA polynucleotides and methods of use

A set of two Class 2 Type II CRISPR polynucleotides forming a dual guide Class 2 Type II polynucleotide, comprising:

• (i) a first Class 2 Type II CRISPR polynucleotide comprising a targeting region comprising ribonucleic acid (RNA) and an activating region adjacent to said targeting region;
• and,
• (ii) a second Class 2 Type II CRISPR polynucleotide comprising an activating region comprising a sequence that is complementary to a sequence in the activating region of the first Class 2 Type II CRISPR polynucleotide,
• wherein the activating region of the first Class 2 Type II CRISPR polynucleotide and the activating region of the second Class 2 Type II CRISPR polynucleotide form a dual guide activating region comprising a mixture of RNA and deoxyribonucleic acid (DNA), wherein the two activating regions are hybridized to each other to form an activating duplex region,
• wherein the activating duplex region comprises a bulge, and the second Class 2 Type II CRISPR polynucleotide further comprises a nexus 3′ to the activating region, and at least one hairpin 3′ to the nexus, and
• wherein the activating duplex region is capable of binding with a Cas9 protein.