Site-specific gene conversion promoter and gene therapeutic
It is intended to provide a preparation whereby an oligonucleotide is efficiently transferred into a cell and the localization thereof in nucleus is promoted, a preparation for facilitating the conversion of the base sequence of a target genomic gene, and a preparation for gene therapy. Namely, a preparation for facilitating site-specific gene conversion which comprises at least a collagen and an oligonucleotide for gene conversion; a preparation for site-specific gene therapy which comprises at least a collagen and an oligonucleotide for gene conversion; a method of arbitrarily converting a specific base on a genomic gene in the nucleus of a cell which comprises bringing the above-described preparation for facilitating gene conversion into contact with the cell; and an preparation for facilitating oligonucleotide intranuclear localization which comprises at least a collagen and an oligonucleoitde.
1 . A preparation for facilitating site-specific gene conversion, comprising at least a collagen and an oligonucleotide for gene conversion.
2 . A preparation for site-specific gene therapy, comprising at least a collagen and an oligonucleotide for gene conversion.
3 . The preparation according to claim 1 , wherein collagen is water-soluble collagen.
4 . The preparation according to claim 3 , wherein the water-soluble collagen is atelocollagen.
5 . The preparation according to any one of claims 1 to 4 , 34 and 35 , wherein the oligonucleotide for gene conversion is an oligonucleotide comprising of at least 20 bases.
6 . The preparation according to claim 1 , wherein the oligonucleotide for gene conversion is a RNA/DNA chimeric oligonucleotide or a DNA oligonucleotide.
7 . The preparation according to claim 5 , wherein the oligonucleotide for gene conversion is an oligonucleotide having a nucleotide sequence forming a Watson-Crick type base pair containing mismatch pairing of 1 to 3 base pairs, with a sense strand or an antisense strand of a gene to be converted.
8 . The preparation according to claim 5 , wherein the oligonucleotide for gene conversion is an oligonucleotide having a nucleotide sequence forming a Watson-Crick type base pair containing deletion or insertion of 1 to 3 bases, with a sense strand or an antisense strand of a gene to be converted.
9 . The preparation according to claim 7 , wherein the mismatch pairing is located at a central part of an oligonucleotide.
10 . The preparation according to claim 8 , wherein the deletion or insertion of bases is located at a central part of an oligonucleotide.
11 . The preparation according to claim 1 or 2 , wherein a dosage form is solution-like.
12 . (canceled)
13 . (canceled)
14 . The preparation according to claim 11 , wherein an oligonucleotide for gene conversion and a collagen form a particulate associated body.
15 . The preparation according to claim 14 , wherein a long diameter of the particulate associated body is 300 nm to 50 μm.
16 . The preparation according to claim 11 , which comprises collagen in a range of 0.01 to 1.0% by weight.
17 . (canceled)
18 . A preparation for facilitating site-specific gene conversion or a preparation for gene therapy, obtained by dissolving collagen in a solution containing 0.01M to 0.1M of a phosphate salt and 0.07M to 0.14M of a sodium salt, adding an oligonucleotide solution for gene conversion containing the same concentration of a phosphate salt and the same concentration of a sodium salt thereto, and stirring this under a temperature of 1 to 10° C.
19 . The preparation according to claim 1 or 2 , wherein a dosage form is solid-like, and an oligonucleotide for gene conversion and a collagen form a particulate associated body.
20 . The preparation according to claim 19 , wherein an oligonucleotide for gene conversion and a collagen form a particulate associated body.
21 . The preparation according to claim 20 , wherein a long diameter of a particulate associated body is 300 nm to 50 μm.
22 . A method of arbitrarily converting a specific base on a genome gene in a nucleus of a cell, which comprises contacting the preparation for facilitating gene conversion as defined in claim 1 or 3 with the cell.
23 . The method according to claim 22 , wherein the cell is a mammal cell.
24 . The method according to claim 22 , wherein the cell is yeast or fungus.
25 . A preparation for facilitating an oligonucleotide intranuclear localization in a nucleus, comprising at least a collagen and an oligonucleotide.
26 . (canceled)
27 . (canceled)
28 . The preparation for facilitating intranuclear localization according to claim 25 , wherein the oligonucleotide and the collagen form a particulate associated body.
29 . The preparation for facilitating intranuclear localization according to claim 28 , wherein a long diameter of the particulate associated body is 300 nm to 50 μm.
30 . The preparation for facilitating intranuclear localization according to claim 25 , which comprises a collagen in a range of 0.01 to 1.0% by weight.
31 . (canceled)
32 . A method of gene conversion of a cell, which comprises contacting a composition comprising at least collagen and an oligonucleotide for gene conversion with a cell in a living body by oral, nasal, via lung, intraportal, intramuscular, subcutaneous, organ surface, intraorgan or transdermal administration.
33 . A method of treating a genetic disease, which comprises converting a gene of a cell of a subject exhibiting a genetic disease by the method of claim 32 .
34 . The preparation according to claim 2 , wherein collagen is water-soluble collagen.
35 . The preparation according to claim 34 , wherein the water-soluble collagen is atelocollagen.