redBS-Seq/caMAB-seq
Reduced Bisulfite Sequencing/5-Carboxylcytosine Methylase-Assisted Bisulfite Sequencing
redBS-Seq relies on the chemical reduction of 5fC to 5hmC by NaBH4. The 5hmC is detected in the same manner as 5mC (Booth et al., 2014). In caMAB-seq, 5fC is first reduced by NaBH4 to 5hmC. Owing to the combination of NaBH4 reduction with M.SssI treatment, 5caC is sequenced as thymidine after bisulfite conversion, whereas 5mC, 5hmC, and 5fC are read as cytosine (Wu et al., 2016)
Advantages:
- Base-resolution approach to map 5fC and 5caC
Disadvantages:
- Requires a high sequencing depth to reduce the false-discovery rate (Neri et al., 2016)
Reagents:
Illumina Library prep and Array Kit Selector
Reviews:
Iatrou A., Kenis G., Rutten B. P., Lunnon K. and van den Hove D. L. Epigenetic dysregulation of brainstem nuclei in the pathogenesis of Alzheimer’s disease: looking in the correct place at the right time? Cell Mol Life Sci. 2017;74:509-523
Neri F., Incarnato D., Krepelova A., Parlato C. and Oliviero S. Methylation-assisted bisulfite sequencing to simultaneously map 5fC and 5caC on a genome-wide scale for DNA demethylation analysis. Nat Protoc. 2016;11:1191-1205
References:
Van Haute L., Dietmann S., Kremer L., et al. Deficient methylation and formylation of mt-tRNA(Met) wobble cytosine in a patient carrying mutations in NSUN3. Nat Commun. 2016;7:12039