Conjugates of guide RNA-Cas protein complex
OwnerMinghong Zhonginventor
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Abstract
Provided herein are compositions of conjugates of a guide RNA(s)-CRISPR Cas protein (RNP) complex. The conjugate comprises a guide RNA(s)-CRISPR Cas protein (RNP) complex and one or more molecules selected from PEG, non-PEG polymers, ligands for cellular receptors, lipids, oligonucleotides, polysaccharides and peptides and chemically linked to the Cas protein and/or guide RNA(s). The conjugates are delivered to targeted cells as RNP complexes, or formed in targeted cells from guide RNA conjugates and an mRNA or a viral vector encoding a Cas protein, or formed in targeted cells from a crRNA conjugates and a viral vector encoding both a Cas protein and a tracrRNA. Also provided are preparation methods and uses of these conjugates.
Claims31
A conjugate comprising:
- i. a clustered regularly interspaced short palindromic repeats (CRISPR)-Cas protein-chemically ligated guide RNA (lgRNA) complex (RNP);
- and
- ii. one or more molecules selected from the group consisting of antibodies, aptamers, and a single strand nucleic acid template,
- wherein said lgRNA comprises:
- (i) a spacer of an oligonucleotide that targets a DNA sequence, and
- (ii) at least one non-nucleotide linker located after its first nucleotide and before its last nucleotide,
- and wherein said one or more molecules are covalently linked to said lgRNA by one or more non-nucleotide linkers at one or more positions other than the 5′-end of said lgRNA.
Pharmaceutical agents comprising:
- i. an lgRNA;
- and
- ii. one or more molecules selected from the group consisting of antibodies, aptamers, and a single strand nucleic acid template,
- wherein said lgRNA is a guide RNA comprising:
- (i) a spacer of an oligonucleotide that targets a DNA sequence;
- and
- (ii) at least one non-nucleotide linker located after its first nucleotide and before its last nucleotide,
- wherein said one or more molecules are covalently linked to said lgRNA by one or more non-nucleotide linkers at one or more positions other than the 5′-end of said LgRNA.
Dependencies are inferred from the claim text.