Crosslinking agent, crosslinking method, method of controlling gene expression, and method of examining gene function
View Patent ↗The present invention provides a crosslinking agent which have photodegradable protective groups at two ends to crosslink double-stranded nucleic acid, a nucleic acid and a protein or a polypeptide, or proteins or polypeptides, in particular, double-stranded RNA; a method for crosslinking a double-stranded RNA or the like using the same; a method for regulating gene expression, which can control the expression of a target gene at an arbitrary timing and location; and a method for examining a gene function. According to the present invention, crosslinking between double-stranded nucleic acids between a nucleic acid and a protein or a polypeptide, or between proteins or polypeptides, in particular, between double-stranded RNA can be easily formed, and in addition, the crosslinking can also be easily removed, so that the expression of a target gene can be easily controlled at an arbitrary timing and location with high efficiency. Hence, as a result, function examination and/or identification of a gene that is expressed at a specific timing and location can be performed. In addition, the RNAi effect of a double-stranded RNA (siRNA) that cannot be easily inhibited by a conventional caged compound can be inhibited, and the expression of a target gene can be easily controlled at an arbitrary timing and location.
1. A compound represented by formula (7):
wherein
Q represents —O—;
X 1 and X 2 each independently represent —H, a hydroxyl group, a substituted alkoxy group, an unsubstituted alkoxy group, a —OC(O)R 11 group, a —NH 3 group, a —NR 11 R 12 group, —R 11 , —F, —Cl, —Br, —I, —COOH, —NO 2 , —C(═O)NHR 11 , —CN, —CHO, —C(═O)R 11 , or —SO 3 H;
Y 1 represents —H, —Cl, —Br, —I, —C(O)OH, —NO 2 , —C(O)NHR 11 , —CN, —C(O)H, —C(O)CH 3 , a benzoxazole-2-yl group, -benzothiazole-2-yl, or -benzoimidazole-2-yl;
Y 2 represents —H, —C(O)OH, or —SO 3 H;
M 1 represents —H;
R 11 and R 12 each independently represent a substituted or an unsubstituted functional group selected from an alkyl group having 1 to 20 carbon atoms, an alkenyl group having 2 to 20 carbon atoms, an alkynyl group having 2 to 20 carbon atoms, an alkoxy group having 1 to 20 carbon atoms, a thioalkoxy group having 1 to 20 carbon atoms, an alkylsulfonyl group having 1 to 20 carbon atoms, an arylsulfonyl group having 4 to 16 carbon atoms, a heteroalkyl group having a total number of carbon and hetero atoms of 2 to 20, a heteroalkenyl group having a total number of carbon and hetero atoms of 2 to 20, a cycloalkyl group having 3 to 8 carbon atoms, a cycloalkenyl group having 3 to 8 carbon atoms, an aryl group having 4 to 16 carbon atoms, a heteroaryl group having a total number of carbon and hetero atoms of 4 to 16, and a heterocyclyl group having a total number of carbon and hetero atoms of 2 to 30;
X 1 and Y 2 may form in combination a group selected from an —O—(CH 2 ) n —O— group, a —C—(CH 2 ) n —O— group, an —O—(CH 2 ) n —C— group, an —O—(CH 2 ) n —N— group, a —N—(CH 2 ) n —O— group, a —N—(CH 2 ) n —N— group, a —C—(CH 2 ) n —N— group, and a —N—(CH 2 ) n —C— group, in which n is 1 or 2;
A 3 and A 4 each independently represent an alkylene group, —O—, NR 1 —, —O—CO—, —CO—O—, —CH 2 —O—CH 2 —, —NR 2 —COO—, —OCO—NR 2 —, —NR 3 —CO—, —CO—NR 3 —, or —O—O—COO—;
R 1 , R 2 and R 3 each independently represent hydrogen or an alkyl group;
T 2 and T 3 each independently represent an alkylene group;
E represents a bond, —NH—, a sulfur atom, an oxygen atom, —O—CO—, or —CO—O—;
p represents an integer of 1 to 6;
each repeat unit of -(T 2 -E)- may be the same or different; and
the compound is a symmetrical compound.
2. A crosslinking agent comprising the compound of claim 1 to crosslink nucleic acids.
3. The compound of claim 1 , wherein -A 3 -(T 2 -E) p -T 3 -A 4 - is 2 Å to 44 Å in length.
4. The compound of claim 1 , wherein -A 3 -(T 2 -E) p -T 3 -A 4 - has a main chain comprising 5 to 80 atoms.
5. The compound of claim 1 , wherein -A 3 -(T 2 -E) p -T 3 -A 4 - is
wherein
m represents an integer of 3 to 78; and
R 1 , R 2 and R 3 each independently represent hydrogen or an alkyl group.
6. The compound of claim 1 , wherein -A 3 -(T 2 -E) p -T 3 -A 4 - is
7. The compound of claim 1 , wherein
X 1 represents —Br;
X 2 represents —H;
Y 1 represents —H; and
Y 2 represents —H.