News

Sahin H, Islam S, Lee AH, Ponzo I, Winiger K, Reichl A, Carell T, Giehr P (2025) Biochemical engineering of 5hmdC-DNA using a Tet3 double-mutant. Comput Struct Biotechnol J, 31:389-396 Link

Abstract:

5-Hydroxymethyl-2'-deoxycytidine (5hmdC) is an important epigenetic marker involved in gene regulation and DNA demethylation. It has potential use as a biomarker for cancer and other diseases due to its significant depletion in various cancers and disease models. This research aimed to develop a reliable and efficient method for generating 5hmdC-containing DNA, addressing limitations in existing techniques. We created a Tet3 stalling mutant that converts 5-methyl-2'-deoxycytidine (5mdC) into a mixture of 5hmdC and 5-formyl-2'-deoxycytidine (5fdC), followed by a reduction step to convert 5fdC to 5hmdC, ensuring a pure 5hmdC state within the CpG context. This method can convert any PCR product, synthetic oligos, and entire genomes into 5hmdC-modified DNA. The principal results demonstrate high specificity and efficiency, providing a robust tool for epigenetic research, cancer diagnostics, and protein binding assays. Additionally, our technique offers 5hmdC-DNA for functional studies and as standards for diagnostic assays.

Read the full paper here: https://www.sciencedirect.com/science/article/pii/S2001037025005586?via%3Dihub