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: Provided is a nonhuman animal model that is obtained by modifying a gene encoding thioredoxin and useful as a disease model of aging, kidney diseases, cardiovascular diseases, hypertension, aortic dissection, chronic obstructive lung disease, age-dependent epilepsy, abnormality of lipid metabolism, anemia, osteoporosis, abnormal immunity, etc. These variety of phenotypes are caused by the fact that a modification of a gene encoding thioredoxin induces hypofunction of thioredoxin expressed in multiple organs throughout the body. The gene encoding thioredoxin is a gene selected from among TXN, TRX, TRX1, RRDX, Txn1, Txn, Trx1 and ADF.

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: Methods are disclosed herein for producing human hepatocytes from human induced pluripotent stem cells. Also provided are transgenic rats for the expansion of human hepatocytes, such as those produced using the methods disclosed herein.

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: Provided is a nonhuman animal model that is obtained by modifying a gene encoding thioredoxin and useful as a disease model of aging, kidney diseases, cardiovascular diseases, hypertension, aortic dissection, chronic obstructive lung disease, age-dependent epilepsy, abnormality of lipid metabolism, anemia, osteoporosis, abnormal immunity, etc. These variety of phenotypes are caused by the fact that a modification of a gene encoding thioredoxin induces hypofunction of thioredoxin expressed in multiple organs throughout the body. The gene encoding thioredoxin is a gene selected from among TXN, TRX, TRX1, RRDX, Txn1, Txn, Trx1 and ADF.

Abstract: An object is to develop a technology enabling utilization, by only an extremely simple technique, of a technology which is widely applicable to mammals without requiring the utilization of an ES cell, and which involves modifying a certain gene by targeting a certain sequence on a genome (genome editing technology based on ZFN or the like). Provided is a technology for efficiently modifying an arbitrary target gene of a mammal, by immersing a pronuclear stage mammalian zygote with an intact zona pellucida into a solution containing a pair of molecules of mRNA having a certain sequence, and performing electroporation treatment through application of multiple square-wave pulses in three steps with the total electric energy of a first electric pulse adjusted within a predetermined range.

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

Abstract: A method for producing a genome edited cell or a non-human organism, comprising the step of introducing (a) an artificial nuclease system which cleaves both ends of a genome editing target region, and (b) a nucleic acid sequence formed by arranging a 5?-side homology arm sequence, a donor DNA sequence, and a 3?-side homology arm sequence in this order from a 5?-side, the 5?-side homology arm sequence being a homologous sequence of one nucleic acid sequence outside the genome editing target region, and the 3?-side homology arm sequence being a homologous sequence of the other nucleic acid sequence outside the genome editing target region, into a cell or a non-human organism.

Abstract: An object is to develop a technology enabling utilization, by only an extremely simple technique, of a technology which is widely applicable to mammals without requiring the utilization of an ES cell, and which involves modifying a certain gene by targeting a certain sequence on a genome (genome editing technology based on ZFN or the like). Provided is a technology for efficiently modifying an arbitrary target gene of a mammal, by immersing a pronuclear stage mammalian zygote with an intact zona pellucida into a solution containing a pair of molecules of mRNA having a certain sequence, and performing electroporation treatment through application of multiple square-wave pulses in three steps with the total electric energy of a first electric pulse adjusted within a predetermined range.

Abstract: Methods are disclosed herein for producing human hepatocytes from human induced pluripotent stem cells. Also provided are transgenic rats for the expansion of human hepatocytes, such as those produced using the methods disclosed herein.

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.

Abstract: An object is to develop a technology enabling utilization, by only an extremely simple technique, of a technology which is widely applicable to mammals without requiring the utilization of an ES cell, and which involves modifying a certain gene by targeting a certain sequence on a genome (genome editing technology based on ZFN or the like). Provided is a technology for efficiently modifying an arbitrary target gene of a mammal, by immersing a pronuclear stage mammalian zygote with an intact zona pellucida into a solution containing a pair of molecules of mRNA having a certain sequence, and performing electroporation treatment through application of multiple square-wave pulses in three steps with the total electric energy of a first electric pulse adjusted within a predetermined range.