5-Formylcytosine (5fC) is a key intermediate in active DNA demethylation, offering a direct snapshot of ongoing epigenetic reprogramming. CD BioSciences provides highly sensitive and specific 5fC analysis services, empowering researchers to map this elusive modification and uncover its role in gene regulation, development, and disease etiology.

Overview of 5-Formylcytosine (5fC)

5-Formylcytosine (5fC) is an oxidative derivative of 5-methylcytosine (5mC), generated primarily by the Ten-Eleven Translocation (TET) family of enzymes during the iterative oxidation process of active DNA demethylation. As a rare and transient epigenetic mark, 5fC is present at low abundance in genomic DNA but is enriched at specific regulatory regions such as enhancers, promoters, and gene bodies of transcriptionally active genes.

Functionally, 5fC is more than just a demethylation intermediate. Emerging evidence suggests it may serve as a stable epigenetic signal in its own right, potentially recruiting specific reader proteins and influencing chromatin structure and transcription. Its unique chemical properties also distinguish it from 5mC and 5hmC. The biological significance of 5fC is profound: its dynamic regulation is crucial for embryonic development, cellular differentiation, and tissue homeostasis. Aberrant 5fC levels or distribution have been implicated in various cancers, neurological disorders, and other diseases, making it a valuable target for understanding epigenetic dysregulation and discovering novel biomarkers.

Fig.1 Active demethylation of 5mC by TET dioxygenases and local enrichment of 5fC. (Kumar D, et al., 2024)

Our Services

CD BioSciences delivers cutting-edge, reliable 5fC analysis services designed to overcome the challenge of detecting this low-abundance modification. We leverage state-of-the-art chemical and sequencing technologies to provide precise, base-resolution mapping and quantitative analysis of 5fC. Our end-to-end solutions, from expert consultation to in-depth bioinformatic interpretation, empower you to elucidate the functional landscape of 5fC in your biological system, accelerating discoveries in developmental biology, neuroepigenetics, and oncology.

Customized 5fC Analysis Solutions

At CD BioSciences, we provide a comprehensive suite of advanced technologies to accurately detect and map 5-formylcytosine (5fC). Our platform integrates highly specific chemical-assisted bisulfite sequencing (e.g., fCAB-seq), selective chemical labeling and enrichment methods, and sensitive mass spectrometry-based quantification. This multi-technology approach enables precise, base-resolution 5fC profiling tailored to diverse sample types and research goals, ensuring you obtain reliable data even from low-abundance epigenetic marks.

1. Global DNA Modification Quantification by LC-MS/MS
2. Global DNA Modification Quantification by ELISA
3. 5fC Chemically Assisted Bisulfite Sequencing (fCAB-seq) Service

1. Chemical-labeling-enabled C-to-T Conversion Sequencing (CLEVER-seq) Service
2. M.SssI Methylase-assisted Bisulfite Sequencing (MAB-seq) Service
3. Other Technologies

Workflow of DNA 5fC Analysis Service

Our 5fC analysis follows a meticulously designed technical pipeline focused on the precise handling of this low-abundance, labile modification. Each step is optimized to ensure maximum specificity, sensitivity, and data integrity, reflecting our deep expertise in advanced epigenomic profiling.

Our Advantages

1. Unmatched Specificity for Rare Modifications: Our platform utilizes rigorously validated chemical and enzymatic strategies designed to exclusively target 5fC, effectively eliminating signal interference from 5mC, 5hmC, and unmodified cytosine.
2. Optimized for Low-Abundance Signal Detection: Proprietary workflow maximizes 5fC recovery and detection sensitivity, ensuring an excellent signal-to-noise ratio even with low-input or complex samples.
3. Quantitative Precision at Single-Base Resolution: We provide both absolute quantification via LC-MS/MS and precise base-resolution mapping via sequencing, enabling confident measurement of global dynamics and locus-specific events.
4. Integrated, Hypothesis-Driven Bioinformatics: Contextual interpretation is provided by integrating 5fC data with gene expression, chromatin states, or public datasets to deliver actionable insights and testable hypotheses.
5. Rigorous, Process-Oriented Quality Assurance: Every project is governed by a stage-gated quality control system. From pre-conversion DNA integrity checks to post-sequencing data validation with orthogonal methods, we ensure reproducibility and data integrity fit for publication and further validation.

Beyond 5fC, CD BioSciences offers a complete portfolio of DNA modification analysis services. We provide tailored, end-to-end solutions for studying 5mC, 5hmC, 5caC, 6mA, 4mC, and 8-oxoG, among others. Our flexible and customizable workflows are designed to support your specific research focus, whether in demethylation pathways, epigenetic inheritance, or disease mechanism exploration. Contact us to build the precise epigenetic profiling strategy your project requires.

Reference
1. Kumar D, Hyder I, Kues W A. 5-Formylcytosine: a new epigenetic player[J]. Signal Transduction and Targeted Therapy, 2024, 9(1): 295.