Protocol for profiling RNA m5C methylation at base resolution using m5C-TAC-seq
Summary: RNA 5-methylcytosine (m5C) is a widespread modification and plays a crucial role in gene expression regulation. Here, we present a protocol for transcriptome-wide m5C methylome profiling at base resolution using bisulfite-free m5C detection strategy enabled by ten-eleven translocation (TET)...
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Elsevier
2025-03-01
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| Series: | STAR Protocols |
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| Online Access: | http://www.sciencedirect.com/science/article/pii/S266616672500005X |
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| author | Xiaoting Zhang Liang Lu Chengqi Yi Xiaoyu Li |
| author_facet | Xiaoting Zhang Liang Lu Chengqi Yi Xiaoyu Li |
| author_sort | Xiaoting Zhang |
| collection | DOAJ |
| description | Summary: RNA 5-methylcytosine (m5C) is a widespread modification and plays a crucial role in gene expression regulation. Here, we present a protocol for transcriptome-wide m5C methylome profiling at base resolution using bisulfite-free m5C detection strategy enabled by ten-eleven translocation (TET)-assisted chemical labeling sequencing (m5C-TAC-seq). We detail steps for TET-assisted chemical labeling, library construction, and data analysis. m5C-TAC-seq enables accurate and robust m5C detection in various RNA species.For complete details on the use and execution of this protocol, please refer to Lu et al.1 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. |
| format | Article |
| id | doaj-art-9682ad26c0074d088cacf901752f9df4 |
| institution | Kabale University |
| issn | 2666-1667 |
| language | English |
| publishDate | 2025-03-01 |
| publisher | Elsevier |
| record_format | Article |
| series | STAR Protocols |
| spelling | doaj-art-9682ad26c0074d088cacf901752f9df42025-02-02T05:29:20ZengElsevierSTAR Protocols2666-16672025-03-0161103599Protocol for profiling RNA m5C methylation at base resolution using m5C-TAC-seqXiaoting Zhang0Liang Lu1Chengqi Yi2Xiaoyu Li3State Key Laboratory of Protein and Plant Gene Research, School of Life Sciences, Peking University, Beijing 100871, ChinaDepartment of Biochemistry and Department of Gastroenterology of the Second Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou 310058, China; Institute of Immunology, Zhejiang University School of Medicine, Hangzhou 310058, ChinaState Key Laboratory of Protein and Plant Gene Research, School of Life Sciences, Peking University, Beijing 100871, China; Peking-Tsinghua Center for Life Sciences, Peking University, Beijing 100871, China; Department of Chemical Biology and Synthetic and Functional Biomolecules Center, College of Chemistry and Molecular Engineering, Peking University, Beijing 100871, China; Corresponding authorDepartment of Biochemistry and Department of Gastroenterology of the Second Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou 310058, China; Corresponding authorSummary: RNA 5-methylcytosine (m5C) is a widespread modification and plays a crucial role in gene expression regulation. Here, we present a protocol for transcriptome-wide m5C methylome profiling at base resolution using bisulfite-free m5C detection strategy enabled by ten-eleven translocation (TET)-assisted chemical labeling sequencing (m5C-TAC-seq). We detail steps for TET-assisted chemical labeling, library construction, and data analysis. m5C-TAC-seq enables accurate and robust m5C detection in various RNA species.For complete details on the use and execution of this protocol, please refer to Lu et al.1 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.http://www.sciencedirect.com/science/article/pii/S266616672500005XBioinformaticsSequence analysisGeneticsGenomicsSequencingRNAseq |
| spellingShingle | Xiaoting Zhang Liang Lu Chengqi Yi Xiaoyu Li Protocol for profiling RNA m5C methylation at base resolution using m5C-TAC-seq STAR Protocols Bioinformatics Sequence analysis Genetics Genomics Sequencing RNAseq |
| title | Protocol for profiling RNA m5C methylation at base resolution using m5C-TAC-seq |
| title_full | Protocol for profiling RNA m5C methylation at base resolution using m5C-TAC-seq |
| title_fullStr | Protocol for profiling RNA m5C methylation at base resolution using m5C-TAC-seq |
| title_full_unstemmed | Protocol for profiling RNA m5C methylation at base resolution using m5C-TAC-seq |
| title_short | Protocol for profiling RNA m5C methylation at base resolution using m5C-TAC-seq |
| title_sort | protocol for profiling rna m5c methylation at base resolution using m5c tac seq |
| topic | Bioinformatics Sequence analysis Genetics Genomics Sequencing RNAseq |
| url | http://www.sciencedirect.com/science/article/pii/S266616672500005X |
| work_keys_str_mv | AT xiaotingzhang protocolforprofilingrnam5cmethylationatbaseresolutionusingm5ctacseq AT lianglu protocolforprofilingrnam5cmethylationatbaseresolutionusingm5ctacseq AT chengqiyi protocolforprofilingrnam5cmethylationatbaseresolutionusingm5ctacseq AT xiaoyuli protocolforprofilingrnam5cmethylationatbaseresolutionusingm5ctacseq |