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AssigneeI'rom Group Co., Ltd.
InventorsKohji KUSANO, Takayuki KITOGO, Makoto INOUE, Tsugumine SHU, Toyotaka MORI
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The present invention provides novel donor polynucleotides formed by linking the two ends of a genomic fragment containing a cleavable site by a polynucleotide carrying a positive selection marker gene and a negative selection marker gene. Use of the donor polynucleotide makes it possible to modify only a target gene with avoiding the possibility of introducing mutations to sequences, called “off-target”, which are other than the target sequence, by introducing cleavage in a homologous site of the donor polynucleotide without introducing cleavage in a target gene locus.
A donor polynucleotide for modifying a genomic sequence, wherein the donor polynucleotide comprises a genomic fragment comprising one or more modifications, wherein both ends of the genomic fragment are linked by a polynucleotide, wherein the linker polynucleotide comprises within it both a positive selection marker gene and a negative selection marker gene, wherein the genomic fragment is cleavable, and wherein the donor polynucleotide may be linear when the cleavable site is severed to become the two ends of the donor polynucleotide chain, or the donor polynucleotide may be circular when the site is connected in the donor polynucleotide.
Dependencies are inferred from the claim text.