Methods and compositions for RNA-guided treatment of HIV infection

A nucleic acid encoding:

• (a) a Clustered Regularly Interspected Short Palindromic Repeat (CRISPR)-associated Cas9 endonuclease;
• (b) two or more guide RNAs (gRNAs), at least two of the two or more gRNAs targeting a cleavage site,
• (i) a first gRNA, the first gRNA being complementary to a first target nucleic acid sequence within a 5′ long terminal repeat (LTR) of a human immunodeficiency virus (HIV) sequence;
• and
• (ii) a second gRNA, the second gRNA being complementary to a second target nucleic acid sequence within a 3′ long terminal repeat (LTR) of the HIV sequence, at least two of the two or more gRNAs being different,
• wherein the nucleic acid is capable of excising the HIV sequence between cleavage sites from a genome in a cell.

A pharmaceutical composition comprising:

• (a) a nucleic acid comprising:
• i. a first nucleic acid sequence encoding a Clustered Regularly Interspaced Short Palindromic Repeat (CRISPR)-associated Cas9 endonuclease;
• ii. a second nucleic acid sequence encoding two or more guide RNAs (gRNAs), at least two of the two or more gRNAs targeting a cleavage site,
• (i) a first gRNA, the first gRNA being complementary to a first target nucleic acid sequence within a 5′ long terminal repeat (LTR) of a human immunodeficiency virus (HIV) sequence;
• and
• (ii) a second gRNA, the second gRNA being complementary to a second target nucleic acid sequence within a 3′ long terminal repeat (LTR) of the HIV sequence, at least two of the two or more gRNAs being different;
• wherein the nucleic acid is capable of excising the HIV sequence between cleavage sites from a genome in a cell;
• and
• (b) a pharmaceutically acceptable excipient.