| 摘要 |
The present invention relates to a 5-formylcytosine specific chemical labeling method and related applications in aspects such as sequencing, detection, imaging, and diagnosis. In the method, a condensation reaction occurs between an active methylene group in an active methylene compound containing a side-chain reactive group and an aldehyde group in 5-formylcytosine or a 1-substituted derivative of 5-formylcytosine, and at the same time an intramolecular reaction occurs between the side-chain reactive group of the active methylene compound and a 4-amino group of cytosine to implement ring closing. By means of the 5-formylcytosine specific chemical labeling method and related compounds of the present invention, detection of the content of 5-formylcytosine in nucleic acid molecules, and specific concentration of 5-formylcytosine-containing nucleic acid samples, and analysis of sequence distribution information of 5-formylcytosine and/or single-base resolution sequence information in nucleic acid molecules and the like may be implemented. The present invention provides various effective research methods in the research fields of epigenetics and nucleic acid biochemistry. |
| 主权项 |
1. A method for specific chemical labeling of 5-formylcytosine or a 1-substituted derivative thereof, comprising the step of reacting an active methylene compound containing a side-chain active group R1—CH2—R2 with the 5-formylcytosine or a 1-substituted derivative thereof, wherein a dehydration condensation reaction occurs between the active methylene compound containing a side-chain active group and a 5-formyl group of cytosine in the 5-formylcytosine or a 1-substituted derivative thereof, and at the same time an intramolecular reaction occurs between the side-chain active group of the active methylene compound and a 4-amino group of cytosine in the 5-formylcytosine or a 1-substituted derivative thereof to implement ring closing, as shown in the equation below: wherein, R represents hydrogen, hydrocarbyl, hydrocarbyl with —OH, —NH2, —CHO and/or —COON, or structures excluding the 5-formylcytosine part of related small molecular compound or polymeric macromolecular compound formed by binding glucosidic bond in ribosyl or deoxyribosyl, 5′- or 3′-phosphate-modified ribosyl or deoxyribosyl, ribonucleic acid or deoxyribonucleic acid to 1-position of the 5-formylcytosine; the hydrocarbyl is C1-C30 linear or branched alkyl, C1-C30 linear or branched alkenyl, or C1-C30 linear or branched alkynyl; R1 is an electrondrawing group selected from cyano, nitro, formyl, carbonyl compound and carboxylic acid and derivatives thereof R2 is an electrondrawing groups selected from cyano, formyl, carbonyl compoundand carboxylic acid and derivatives thereof R3 is an unsubstituted C1-C30 linear or branched alkyl, alkenyl or alkynyl, or a C1-C30 linear or branched alkyl, alkenyl or alkynyl substituted with —OH, —NH2, —CHO, —COON and/or azido, or biotin; and R1 and R2 are independent from each other or forming a ring directly by bonding with each other or forming a ring indirectly by bonding via an atom C, Nor O. |
