Figure 1.
The structures of the cytosine derivatives.
5-mC modification is catalyzed by DNA methyltransferase (DNMT). 5-mC can be demethylated through the oxidation of 5-mC by TET proteins to produce 5-hmC, 5-fC and 5-caC. TDG, thymine-DNA glycosylase; BER, base-excision repair.
Table 1.
The sequences of the 27mer cytosine derivative-containing and the control ODNs used for replication studies.
Figure 2.
A schematic diagram outlining the experimental procedures.
The 27mer cytosine derivative-containing ODNs with barcodes were ligated to the EcoR I-linearized M13 vector, mixed at equal amounts and subjected to in-vivo replication. The harvested M13 progenies were amplified with barcoded PCR primers, and equal amounts of PCR products from different cell lines were mixed and subjected to NGS library construction and sequencing.
Figure 3.
Bypass efficiencies and mutation frequencies of cytosine derivatives.
(A) Bypass efficiencies of 5-hmC, 5-foC and 5-caC. (B–E) Mutation frequencies of control (B), 5-hmC (C), 5-foC (D) and 5-caC (E). The data represent the means and standard deviations of results from three independent experiments.