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• W4311577892 abstract "Abstract Background Chromatin-bound RNAs are the primary product of transcription that undergo on-chromatin processing such as capping, splicing, and polyadenylation. These processing steps then determine the fate of RNAs. Albeit its vital importance, a simple and robust method for isolating different fractions of chromatin-bound RNAs is missing in plants. Result Here, we describe our updated method and the associated step-by-step protocol for chromatin-bound RNAs isolation in A. thaliana . The chromatin-bound RNAs isolation is based on the 1 M UREA wash that removes the majority of non-chromatin-associated proteins from the nucleus, as previously developed in mammalian cells. On-demand, the isolated chromatin-bound RNAs can be either used directly for gene-specific analysis or subject to further rRNA removal and also the optional polyadenylated RNA removal, followed by high-throughput sequencing. Detailed protocols for these procedures are also provided. Comparison of sequencing results of chromatin-bound RNAs with and without polyadenylated RNA removal revealed that a small fraction of CB-RNAs is polyadenylated but not yet fully spliced, representing RNA-processing intermediate on-chromatin. Conclusion This optimized chromatin-bound RNAs purification method is simple and robust and can be used to study transcription and its-coupled RNA processing in plants." @default.
• W4311577892 created "2022-12-27" @default.
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• W4311577892 date "2022-12-12" @default.
• W4311577892 modified "2023-10-14" @default.
• W4311577892 title "A simple and robust method for isolating and analyzing chromatin-bound RNAs in Arabidopsis" @default.
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• W4311577892 doi "https://doi.org/10.1186/s13007-022-00967-y" @default.
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