journal article·2024-10-08

Multiplexed In Situ Imaging of Site-Specific m6A Methylation with Proximity Hybridization Followed by Primer Exchange Amplification (m6A-PHPEA)

Minghui Song, Junyan Wang, Jianing Hou, Ting Fu, Yawei Feng, Wenyi Lv, Feng Ge, Ruizi Peng, Da Han, Weihong Tan

ACS Nano· Volume 18· Issue 40
Publisher page

Abstract

Post-transcriptional modification of N6-methyladenosine (m6A) is crucial for ribonucleic acid (RNA) metabolism and cellular function. The ability to visualize site-specific m6A methylation at the single-cell level would markedly enhance our understanding of its pivotal regulatory functions in the field of epitranscriptomics. Despite this, current in situ imaging techniques for site-specific m6A are constrained, posing a significant barrier to epitranscriptomic studies and pathological diagnostics. Capitalizing on the precise targeting capability of deoxyribonucleic acid (DNA) hybridization and the high specificity of the m6A antibody, we present a method, termed proximity hybridization followed by primer exchange amplification (m6A-PHPEA), for the site-specific imaging of m6A methylation within cells. This approach enables high-resolution, single-cell imaging of m6A methylation across various RNA molecules coupled with efficient signal amplification. We successfully imaged three distinct m6A methylation sites concurrently in multiple cell types, revealing cell-to-cell variability in expression levels. This method promises to illuminate the dynamics of m6A-modified RNAs, potentially revolutionizing epitranscriptomic research and the development of advanced pathological diagnosis for chemical modifications.
Source: openalex_inverted_index · Quality: baseline

Research context

OpenAlex topics: RNA modifications and cancer · HVDC Systems and Fault Protection · Cardiac Structural Anomalies and Repair

SciVX discovery tags are terminology-based. OpenAlex topics are source-provided classifications.

Affiliations

The Key Laboratory of Zhejiang Province for Aptamers and Theranostics, Zhejiang Cancer Hospital, Hangzhou Institute of Medicine (HIM), Chinese Academy of Sciences, Hangzhou, Zhejiang 310022, P. R. China

Hangzhou Institute for Advanced Study, University of Chinese Academy of Sciences, Hangzhou, Zhejiang 310024, P. R. China

Institute of Chemistry, Chinese Academy of Sciences (CAS), Beijing 100190, P. R. China

Institute of Molecular Medicine (IMM), Renji Hospital, Shanghai Jiao Tong University School of Medicine, and College of Chemistry and Chemical Engineering, Shanghai Jiao Tong University, Shanghai 200240, P. R. China

Molecular Science and Biomedicine Laboratory (MBL), State Key Laboratory of Chemo/Biosensing and Chemometrics, College of Chemistry and Chemical Engineering, College of Biology, Aptamer Engineering Center of Hunan Province, Hunan University, Changsha, Hunan 410082, P. R. China

Funding

Natural Science Foundation of Hunan Province · China Postdoctoral Science Foundation · National Natural Science Foundation of China · Natural Science Foundation of Zhejiang Province · Zhejiang Provincial Jianbing Lingyan Key Research and Development Project · Zhejiang Leading Innovation and Entrepreneurship Team

References (43)

Show source-provided references
1. 10.1016/j.bbadis.2022.16651210.1016/j.bbadis.2022.166512 ↗
2. 10.1038/hdy.2010.5410.1038/hdy.2010.54 ↗
3. 10.1038/s41576-022-00468-710.1038/s41576-022-00468-7 ↗
4. 10.1038/nchembio.48210.1038/nchembio.482 ↗
5. 10.1093/nar/gkab108310.1093/nar/gkab1083 ↗
6. 10.1016/j.cell.2014.08.00510.1016/j.cell.2014.08.005 ↗
7. 10.1038/s41580-019-0168-510.1038/s41580-019-0168-5 ↗
8. 10.1038/nature1273010.1038/nature12730 ↗
9. 10.1016/j.ccell.2017.02.01310.1016/j.ccell.2017.02.013 ↗
10. 10.1038/s41571-023-00774-x10.1038/s41571-023-00774-x ↗
11. 10.1038/nature1423410.1038/nature14234 ↗
12. 10.1016/j.tig.2022.10.00410.1016/j.tig.2022.10.004 ↗
13. 10.1186/s12943-019-1082-310.1186/s12943-019-1082-3 ↗
14. 10.1016/j.ccell.2020.02.00410.1016/j.ccell.2020.02.004 ↗
15. 10.1186/s13578-020-00502-310.1186/s13578-020-00502-3 ↗
16. 10.15252/embj.202010597710.15252/embj.2020105977 ↗
17. 10.1038/nature1111210.1038/nature11112 ↗
18. 10.1038/s41587-022-01487-910.1038/s41587-022-01487-9 ↗
19. 10.1038/s41592-019-0570-010.1038/s41592-019-0570-0 ↗
20. 10.1038/s41589-020-0525-x10.1038/s41589-020-0525-x ↗
21. 10.1038/s41587-022-01587-610.1038/s41587-022-01587-6 ↗
22. 10.1093/nar/gkad20010.1093/nar/gkad200 ↗
23. 10.1002/anie.20200726610.1002/anie.202007266 ↗
24. 10.1002/anie.20180794210.1002/anie.201807942 ↗
25. 10.1261/rna.041178.11310.1261/rna.041178.113 ↗
26. Zhu Y. · Interdiscip. Med. · 2023 · 2 · e2023004310.1002/INMD.20230043 ↗
27. 10.1002/agt2.1610.1002/agt2.16 ↗
28. 10.1093/nar/gkad78710.1093/nar/gkad787 ↗
29. 10.1002/anie.20210911810.1002/anie.202109118 ↗
30. 10.1038/nchem.287210.1038/nchem.2872 ↗
31. 10.1021/jacs.9b0547010.1021/jacs.9b05470 ↗
32. 10.1002/anie.20201119810.1002/anie.202011198 ↗
33. 10.1038/s41592-019-0404-010.1038/s41592-019-0404-0 ↗
34. 10.1038/s41587-019-0207-y10.1038/s41587-019-0207-y ↗
35. 10.1016/j.molcel.2016.03.02110.1016/j.molcel.2016.03.021 ↗
36. 10.1038/s41419-017-0129-x10.1038/s41419-017-0129-x ↗
37. 10.1093/nar/gku127610.1093/nar/gku1276 ↗
38. 10.1021/jacs.2c0103610.1021/jacs.2c01036 ↗
39. 10.1126/science.aaa609010.1126/science.aaa6090 ↗
40. 10.1039/C6SC05420J10.1039/C6SC05420J ↗
41. 10.1038/s41592-022-01604-110.1038/s41592-022-01604-1 ↗
42. 10.1038/nmeth.283510.1038/nmeth.2835 ↗
43. 10.1002/anie.20161172910.1002/anie.201611729 ↗