Deaminase-assisted method for single-base-resolution localization analysis of 5-carboxycytosine modifications in DNA

A carboxycytosine and analysis method technology, applied in the field of 5-carboxycytosine modified single base resolution positioning analysis, can solve the problems of increasing purification steps, DNA loss, etc., achieve high deamination efficiency, easy operation, and facilitate positioning The effect of the analysis
CN110358817BActive Publication Date: 2021-03-16WUHAN UNIV
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Patent Information

Authority / Receiving Office
CN · China
Patent Type
Patents(China)
Current Assignee / Owner
WUHAN UNIV
Publication Date
2021-03-16

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Abstract

The invention discloses a single-base resolution localization analysis method for 5-carboxycytosine modification in DNA assisted by deaminase. The cytosine deaminase APOBEC3A protein is used to efficiently deaminate normal cytosine, 5-methylcytosine, 5-hydroxymethylcytosine and 5-aldehyde cytosine in DNA, except 5-carboxycytosine. Then PCR amplification is carried out, followed by sequencing to obtain the site information of 5-carboxycytosine. This method has the advantages of high sensitivity, high selectivity and simple operation, and can be adopted to directly obtain the single base resolution location of 5-carboxycytosine in DNA without the treatment of a DNA sample with bisulfite.
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Description

technical field

[0001] The invention relates to the field of biotechnology, in particular to a deaminase-assisted single-base resolution positioning analysis method for 5-carboxycytosine modification in DNA. Background technique

[0002] In mammalian DNA, the 5-position of cytosine will undergo methylation modification under the action of DNA methyltransferase to form 5-methylcytosine (5-mC). 5-mC participates in many important biological processes, such as genome imprinting, gene expression regulation, etc., and is an important epigenetic modification. 5-methylcytosine can be further oxidized successively by TET proteins to 5-hydroxymethylcytosine (5-hmC), 5-formylcytosine (5-foC) and 5-carboxycytosine (5-caC) , forming a new modification. These newly discovered modifiers are closely related to a variety of important physiological functions, such as cell differentiation, cell reprogramming, nervous system development, the occurrence and development of diseases, and so on....

Claims

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