State Key Laboratory of Protein and Plant Gene Research, School of Life Sciences , Peking University , Beijing 100871 , China.
Peking-Tsinghua Center for Life Sciences , Academy for Advanced Interdisciplinary Studies, Peking University , Beijing 100871 , China.
J Am Chem Soc. 2018 Oct 17;140(41):13190-13194. doi: 10.1021/jacs.8b08297. Epub 2018 Oct 4.
High-resolution detection of genome-wide 5-hydroxymethylcytosine (5hmC) sites of small-scale samples remains challenging. Here, we present hmC-CATCH, a bisulfite-free, base-resolution method for the genome-wide detection of 5hmC. hmC-CATCH is based on selective 5hmC oxidation, chemical labeling and subsequent C-to-T transition during PCR. Requiring only nanoscale input genomic DNA samples, hmC-CATCH enabled us to detect genome-wide hydroxymethylome of human embryonic stem cells in a cost-effective manner. Further application of hmC-CATCH to cell-free DNA (cfDNA) of healthy donors and cancer patients revealed base-resolution hydroxymethylome in the human cfDNA for the first time. We anticipate that our chemical biology approach will find broad applications in hydroxymethylome analysis of limited biological and clinical samples.
高分辨率检测小样本范围的全基因组 5-羟甲基胞嘧啶（5hmC）位点仍然具有挑战性。在这里，我们提出了 hmC-CATCH，这是一种无需亚硫酸氢盐、基于碱基分辨率的全基因组 5hmC 检测方法。hmC-CATCH 基于选择性 5hmC 氧化、化学标记和随后 PCR 过程中的 C 到 T 转换。hmC-CATCH 仅需要纳米级输入的基因组 DNA 样本，使我们能够以具有成本效益的方式检测人类胚胎干细胞的全基因组羟甲基组。进一步将 hmC-CATCH 应用于健康供体和癌症患者的无细胞 DNA（cfDNA），首次在人类 cfDNA 中揭示了基于碱基分辨率的羟甲基组。我们预计，我们的化学生物学方法将在有限的生物和临床样本的羟甲基组分析中得到广泛应用。
