Patent ID: 11920152
Assignee: THE GENERAL HOSPITAL CORPORATION
Field: Biotechnology (Chemistry)
Classification: CPC C | IPC C

Claim 0:
1. A method of inducing a single or double-stranded break in a target region of a double-stranded DNA molecule the method comprising expressing in or introducing into the cell:
a Cas9 nuclease or nickase;
a tracrRNA; and
a crRNA that includes one or more nucleotides that are modified,
wherein the ribose phosphate backbone has been replaced by a polyamide chain,
wherein the crRNA consists of the sequence:, (SEQ ID NO: 5)

(X17-20)GUUUUAGAGCUA;

(SEQ ID NO: 6)

(X17-20)GUUUUAGAGCUAUGCUGUUUUG; or

(SEQ ID NO: 7)

(X17-20)GUUUUAGAGCUAUGCU,

wherein X17-20 is a sequence of 17-20 nucleotides that are complementary to the target sequence, preferably the target sequence immediately 5′ of a protospacer adjacent motif (PAM).