Abstract: The present inventors have found that by cultivating insect cells, into which the Cas3 gene has been introduced, at relatively low temperatures, it is possible to efficiently express recombinant Cas3 proteins with maintained activity, and by purifying the soluble fractions of these cells, it is possible to collect active forms of the recombinant Cas3 proteins in high purity and high yield.

Abstract: A CRISPR-Cas3 system was successfully established in a eukaryotic cell.

Abstract: A CRISPR-Cas3 system was successfully established in a eukaryotic cell.

Abstract: A CRISPR-Cas3 system was successfully established in a eukaryotic cell.

Abstract: It has been found that by mixing a single-stranded probe DNA whose cleavage can be detected in a reaction system containing a CRISPR-Cas3 system and a sample from which to detect a target DNA, it is possible to detect the target DNA in the sample by using, as indication, a signal generated by the cleavage of the single-stranded probe DNA.

Abstract: The present inventors have found that the use of a site-specific nuclease system which includes a combination of a molecule that simultaneously targets and cleaves one homology arm sequence of the donor DNA and the genomic sequence corresponding to the homology arm sequence, and a molecule that targets and cleaves the genomic region in the vicinity of the cleavage site by that molecule causes repair between genomic DNA and donor DNA through both non-homologous end joining and homologous recombination, making it possible to produce cells and organisms with knocked-in long donor sequences with high efficiency and accuracy.

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: A CRISPR-Cas3 system was successfully established in a eukaryotic cell.

Abstract: This invention provides a method for knock-in of a donor DNA into the genome of a cell, comprising introducing at least one artificial nuclease system capable of cleaving target sequence(s) of the cell genome, the donor DNA, and two single-stranded oligonucleotides (ssODNs) into the cell, the artificial nuclease system cleaving the target sequence(s) on the cell genome, the two ssODNs each complementary to one of the ends generated by the target sequence cleavage in the cell genome and to one of the introduction ends of the donor DNA, the donor DNA being knocked-in at the cleavage site via the two ssODNs.

Abstract: This invention provides a method for knock-in of a donor DNA into the genome of a cell, comprising introducing at least one artificial nuclease system capable of cleaving target sequence(s) of the cell genome, the donor DNA, and two single-stranded oligonucleotides (ssODNs) into the cell, the artificial nuclease system cleaving the target sequence(s) on the cell genome, the two ssODNs each complementary to one of the ends generated by the target sequence cleavage in the cell genome and to one of the introduction ends of the donor DNA, the donor DNA being knocked-in at the cleavage site via the two ssODNs.