| Title |
A bisulfite-free approach for base-resolution analysis of genomic 5-carboxylcytosine / |
| Authors |
Ličytė, Janina ; Gibas, Povilas ; Skardžiūtė, Kotryna ; Stankevičius, Vaidotas ; Rukšėnaitė, Audronė ; Kriukienė, Edita |
| DOI |
10.1016/j.celrep.2020.108155 |
| Full Text |
|
| Is Part of |
Cell reports.. Cambridge : Cell Press. 2020, vol. 32, iss. 11, art. no. 108155, p. [1-21].. eISSN 2211-1247 |
| Keywords [eng] |
5-carboxylcytosine ; 5-methylcytosine ; 5caC ; 5mC ; C-C bond cleavage ; covalent labeling ; demethylation ; mouse ESCs ; pluripotency states. |
| Abstract [eng] |
Due to an extreme rarity of 5-carboxylcytosine (5caC) in the mammalian genome, investigation of its role brings a considerable challenge. Methods based on bisulfite sequencing have been proposed for genome-wide 5caC analysis. However, bisulfite-based sequencing of scarcely abundant 5caC demands significant experimental and computational resources, increasing sequencing cost. Here, we present a bisulfite-free approach, caCLEAR, for high-resolution mapping of 5caCGs. The method uses an atypical activity of the methyltransferase eM.SssI to remove a carboxyl group from 5caC, generating unmodified CGs, which are localized by uTOP-seq sequencing. Validation of caCLEAR on model DNA systems and mouse ESCs supports the suitability of caCLEAR for analysis of 5caCGs. The 5caCG profiles of naive and primed pluripotent ESCs reflect their distinct demethylation dynamics and demonstrate an association of 5caC with gene expression. Generally, we demonstrate that caCLEAR is a robust economical approach that could help provide deeper insights into biological roles of 5caC. |
| Published |
Cambridge : Cell Press |
| Type |
Journal article |
| Language |
English |
| Publication date |
2020 |
| CC license |
|
