Abstract: A method of skipping a target exon of a gene of interest in a genome uses CRISPR-Cas and guide RNA. The guide RNA contains a spacer sequence such that the site of cleavage by the CRISPR-Cas is positioned within 80 bases from the splice donor site immediately before the target exon or the splice acceptor site immediately after the target exon.

Abstract: A production method for a genetically modified megakaryocyte, the method including a step of introducing a CRISPR-associated (Cas) family protein and guide RNA (gRNA) into a megakaryocyte to modify a subject gene, in which the Cas family protein and the gRNA to be introduced into the megakaryocyte in the step form a complex beforehand. This production method is useful as a technique for producing a genetically modified megakaryocyte and a modified platelet.

Abstract: The present invention provides a means excellent in efficiency of introduction of a nucleic acid, a protein, a complex thereof, or the like into cells, in particular a method using a solution for transduction.

Abstract: A production method of genomic DNA in which a deletion of more than 100 bases of nucleotides is introduced into a target region of the genomic DNA, the method includes a contact step of bringing a type I CRISPR associated complex for anti-viral defense (type I Cascade complex), CRISPR RNA (crRNA), and Cas3 protein into contact with the genomic DNA.

Abstract: The present invention provides a delivery technique for delivering a gene modification tool capable of providing a high gene modification efficiency in cells. The composition according to the present invention is a composition for inducing gene modification at a target gene locus in a cell, the composition containing 1) a compound represented by formula (I) or a salt thereof; 2) a structural lipid; and 3) a guide RNA or a DNA including a sequence encoding the guide RNA, and/or an RNA-guided nuclease or a nucleic acid including a sequence encoding the RNA-guided nuclease. In formula (I), n represents an integer of 2 to 5, R represents a linear C1-5 alkyl group, a linear C7-11 alkenyl group, or a linear C11 alkadienyl group, and wavy lines each independently represent a cis-type bond or a trans-type bond.

Abstract: A virus-like particle encapsulating a target protein is provided. The virus-like particle contain a Gag protein, and the Gag protein forms a dimer with the target protein.

Abstract: A method of skipping a target exon of a gene of interest in a genome uses CRISPR-Cas and guide RNA. The guide RNA contains a spacer sequence such that the site of cleavage by the CRISPR-Cas is positioned within 80 bases from the splice donor site immediately before the target exon or the splice acceptor site immediately after the target exon.

Abstract: A method for producing, from a donor cell, a low-antigenic cell in which a rejection reaction is reduced in a case where the cell is allogeneically transplanted into a recipient, the method including: determining human leukocyte antigen (HLA) alleles for the donor cell and the recipient, respectively; specifying an HLA allele that is present in the donor cell but is not present in the recipient; and disrupting or modifying the specified HLA allele to obtain a cell population including a cell not expressing an HLA protein specific to the donor cell, in which the cell not expressing the HLA protein specific to the donor cell is the low-antigenic cell.

Abstract: Provided is a method for inducing T cells for a cell-based immunotherapy, comprising the steps of: (1) providing Rag 1 and/or Rag 2 gene knockout human pluripotent stem cells bearing genes encoding a T cell receptor specific for a desired antigen, and (2) inducing T cells from the pluripotent stem cells of step (1). Further provided are a cell-based immunotherapy method that uses the T cells for the cell-based immunotherapy and an iPS cell bank for the cell-based immunotherapy.

Abstract: A stem cell expression cassette, comprising a nucleic acid comprising a pluripotent stem cell specific promoter, and a tag sequence, wherein the pluripotent stem cell specific promoter and tag sequences are operatively linked, is provided. Also provided are methods of identifying and methods of selecting a pluripotent cell, using the stem cell expression cassette.