| Autor: |
Liyao Mai, Zebin Wen, Yulong Zhang, Yu Gao, Guanchuan Lin, Zhiwei Lian, Xiang Yang, Jingjing Zhou, Xianwei Lin, Chaochao Luo, Wanwan Peng, Caiming Chen, Duolian Liu, Junxiao Zhang, Sadie L. Marjani, Qian Tao, Xuedong Wu, Sherman M. Weissman, Xinghua Pan |
| Rok vydání: |
2023 |
| DOI: |
10.1101/2023.05.22.541674 |
| Popis: |
DNA methylation is essential for a wide variety of biological processes, yet the development of a highly efficient and robust technology remains a challenge for routine single-cell analysis. We developed a multiplex scalable single-cell reduced representation bisulfite sequencing (msRRBS) technology with off-the-shelf reagents and equipment. It allows cell-specific barcoded DNA fragments of individual cells to be pooled before bisulfite conversion, free of enzymatic modification or physical capture of the DNA ends, and achieves unparalleled read mapping rates of 62.51%, covering 59.95% of CpG islands and 71.62% of promoters in K562 cells on average. Its reproducibility is shown in duplicates of bulk cells with near perfect correlation (R=0.97-99). At a low 1 Mb of clean reads, msRRBS provides consistent coverage of CpG islands and promoters, outperforming the conventional methods with orders of magnitude reduction in cost. Here, we use this method to characterize the distinct methylation patterns and cellular heterogeneity of 6 cell lines, and leukemia and hepatocellular carcinoma models. Taking 4 hours of hands-on time, msRRBS offers a unique, highly efficient approach for dissecting methylation heterogeneity in a variety of multicellular systems. |
| Databáze: |
OpenAIRE |
| Externí odkaz: |
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