Abstract: A polynucleotide, comprising the following base sequences: (a) a base sequence encoding a fusion protein of a nuclease-deficient CRISPR effector protein and a transcription repressor, and (b) a base sequence encoding a guide RNA targeting a continuous region set forth in SEQ ID NO: 2, 3, 4, 20, 51, 68, 144, 148, 152, 162, 164, or 167 in the expression regulatory region of human DUX4 gene is expected to be useful for treating or preventing facioscapulohumeral muscular dystrophy (FSHD).

Abstract: A mutant SaCas9 protein such as a protein having an amino acid sequence resulting from mutations of glutamic acid at the 782-position to lysine (E782K), leucine at the 800-position to arginine (L800R), asparagine at the 968-position to arginine (N968R), asparagine at the 985-position to alanine (N985A), arginine at the 991-position to alanine (R991A), alanine at the 1021-position to serine (A1021S), threonine at the 927-position to lysine (T927K), lysine at the 929-position to asparagine (K929N), and isoleucine at the 1017-position to phenylalanine (I1017F) in SEQ ID NO: 2 has relaxed restriction on target sequence while maintaining binding ability to guide RNA, and is useful as a tool for gene editing.

Abstract: The present invention aims to provide a novel therapeutic approach to human muscular dystrophy (particularly MDC1A). The present invention provide a polynucleotide comprising the following base sequences: (a) a base sequence encoding a fusion protein of a nuclease-deficient CRISPR effector protein and a transcription activator, and (b) abase sequence encoding (i) a guide RNA targeting continuous region set forth in SEQ ID NO: 15, 20, 25, 50, 56, or 61, (ii) a guide RNA targeting a continuous region set forth in SEQ ID NO: 124, or (iii) a guide RNA targeting a continuous region set forth in SEQ ID NO: 178, 193, or 195, in the expression regulatory region of human LAMA1 gene.

Abstract: The present invention provides a method of treating a disease associated with elevated KRAS activity or expression in a subject, comprising suppressing KRAS expression in the subject by targeting an expression regulatory region of KRAS gene using a CRISPR-Guide Nucleotide Directed Modulation (GNDM). Also, provided is a CRISPR-GNDM system for suppressing KRAS expression comprising (a) a protein selected from the group consisting of dCas9 or dCpf1, a fusion protein of dCas9 or dCpf1 and Kruppel associated box (KRAB), and (b) a guide nucleotide targeting an expression regulatory region of KRAS gene.

Abstract: A mutant SaCas9 protein such as a protein having an amino acid sequence resulting from mutations of glutamic acid at the 782-position to lysine (E782K), leucine at the 800-position to arginine (L800R), asparagine at the 968-position to arginine (N968R), asparagine at the 985-position to alanine (N985A), arginine at the 991-position to alanine (R991A), alanine at the 1021-position to serine (A1021S), threonine at the 927-position to lysine (T927K), lysine at the 929-position to asparagine (K929N), and isoleucine at the 1017-position to phenylalanine (I1017F) in SEQ ID NO: 2 has relaxed restriction on target sequence while maintaining binding ability to guide RNA, and is useful as a tool for gene editing.

Abstract: Polynucleotides comprising the following base sequences: (a) a base sequence encoding a fusion protein of a nuclease-deficient CRISPR effector protein and a transcription activator, and (b) a base sequence encoding a guide RNA targeting a continuous region of 18 to 24 nucleotides in length in a region set forth in SEQ ID NO: 104, 105, 135, 141, 153, 167, or 172 in the expression regulatory region of human Utrophin gene are expected to be useful for treating or preventing DUCHENNE muscular dystrophy or BECKER muscular dystrophy.

Abstract: The present invention provides a method of treating a disease associated with activated MYD88 signaling in a subject, including suppressing MYD88 expression in a subject by targeting an expression regulatory region of MYD88 gene by using a CRISPR-Guide Nucleotide Directed Modulation (GNDM), without affecting the expression of an adjacent ACAA1 gene. Also, provided is a CRISPR-GNDM system for suppressing MYD88, including expression comprising (a) a protein selected from the group consisting of dCas9 or dCpf1, a fusion protein of dCas9 or dCpf1 and Kruppel associated box (KRAB) and (b) a guide RNA (gRNA) targeting an expression regulatory region of MYD88 gene.

Abstract: A protein having a binding ability to guide RNA and consisting of a sequence containing an amino acid sequence wherein a continuous deletion region is present between the 481-position and the 649-position in the amino acid sequence shown in SEQ ID NO: 2, the deletion region containing (i) all or a part of L1 domain (481- to 519-positions), and (ii) entire HNH domain (520- to 628-positions), and further optionally containing (iii) all or a part of L2 domain (629- to 649-positions), wherein amino acids adjacent to each of the deletion region are linked by a linker consisting of 3 to 10 amino acid residues functions as a miniaturized dSaCas9 protein while maintaining DNA binding affinity. Use of the miniaturized dSaCas9 protein makes it possible to mount many genes into vectors.

Abstract: Polynucleotides comprising the following base sequences: (a) a base sequence encoding a fusion protein of a nuclease-deficient CRISPR effector protein and a transcription activator, and (b) a base sequence encoding a guide RNA targeting a continuous region of 18 to 24 nucleotides in length in a region set forth in SEQ ID NO: 104, 105, 135, 141, 153, 167, or 172 in the expression regulatory region of human Utrophin gene are expected to be useful for treating or preventing DUCHENNE muscular dystrophy or BECKER muscular dystrophy.

Abstract: The present invention provides a transcription activator consisting of not more than 200 amino acid sequences and containing VP64 and a transcription activation site of RTA. The present invention also provides a complex of a nucleic acid sequence-recognizing module specifically binding to a target nucleotide sequence in a double-stranded DNA and the transcription activator.

Abstract: The present invention provides a method of treating a disease associated with activated MYD88 signaling in a subject, including suppressing MYD88 expression in a subject by targeting an expression regulatory region of MYD88 gene by using a CRISPR-Guide Nucleotide Directed Modulation (GNDM), without affecting the expression of an adjacent ACAA1 gene. Also, provided is a CRISPR-GNDM system for suppressing MYD88, including expression comprising (a) a protein selected from the group consisting of dCas9 or dCpf1, a fusion protein of dCas9 or dCpf1 and Kruppel associated box (KRAB) and (b) a guide RNA (gRNA) targeting an expression regulatory region of MYD88 gene.

Abstract: The present invention relates to a protein having a binding ability to guide RNA and consisting of a sequence comprising an amino acid sequence wherein a continuous deletion region is present between the 721-position and the 755-position in the amino acid sequence shown in SEQ ID NO: 2, wherein amino acids adjacent to each of the deletion region are linked by a linker consisting of 3 to 10 amino acid residues. The binding ability to the guide RNA, and further, the DNA binding affinity, are maintained even though the protein has a deletion region and is smaller than the full-length dSaCas9. Use of the miniaturized dSaCas9 protein makes it possible to mount many genes into vectors.

Abstract: A mutant SaCas9 protein such as a protein having an amino acid sequence resulting from mutations of glutamic acid at the 782-position to lysine (E782K), leucine at the 800-position to arginine (L800R), asparagine at the 968-position to arginine (N968R), asparagine at the 985-position to alanine (N985A), arginine at the 991-position to alanine (R991A), alanine at the 1021-position to serine (A1021S), threonine at the 927-position to lysine (T927K), lysine at the 929-position to asparagine (K929N), and isoleucine at the 1017-position to phenylalanine (I1017F) in SEQ ID NO: 2 has relaxed restriction on target sequence while maintaining binding ability to guide RNA, and is useful as a tool for gene editing.

Abstract: The present invention provides a method of treating a disease associated with elevated KRAS activity or expression in a subject, comprising suppressing KRAS expression in the subject by targeting an expression regulatory region of KRAS gene using a CRISPR-Guide Nucleotide Directed Modulation (GNDM). Also, provided is a CRISPR-GNDM system for suppressing KRAS expression comprising (a) a protein selected from the group consisting of dCas9 or dCpf1, a fusion protein of dCas9 or dCpf1 and Kruppel associated box (KRAB), and (b) a guide nucleotide targeting an expression regulatory region of KRAS gene.