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Exploring RNA transcription and turnover in vivo by using click chemistry
Open accessAbstract
We describe a chemical method to detect RNA synthesis in cells, based on the biosynthetic incorporation of the uridine analog 5-ethynyluridine (EU) into newly transcribed RNA, on average once every 35 uridine residues in total RNA. EU-labeled cellular RNA is detected quickly and with high sensitivity by using a copper (I)-catalyzed cycloaddition reaction (often referred to as “click” chemistry) with fluorescent azides, followed by microscopic imaging. We demonstrate the use of this method in cultured cells, in which we examine the turnover of bulk RNA after EU pulses of varying lengths. We also use EU to assay transcription rates of various tissues in whole animals, both on sections and by whole-mount staining. We find that total transcription rates vary greatly among different tissues and among different cell types within organs.
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Cho, W.-K., Spille, J.-H., Hecht, M., Lee, C., Li, C., Grube, V., & Cisse, I. I. (2018) · Science · doi:10.1126/science.aar4199 · Cited by 1,403
SARS-CoV-2 Disrupts Splicing, Translation, and Protein Trafficking to Suppress Host Defenses
Banerjee, A. K., Blanco, M. R., Bruce, E. A., Honson, D. D., Chen, L. M., Chow, A., Bhat, P., Ollikainen, N., Quinodoz, S. A., Loney, C., et al. (2020) · Cell · doi:10.1016/j.cell.2020.10.004 · Cited by 583
Yurdagul, A., Subramanian, M., Wang, X., Crown, S. B., Ilkayeva, O. R., Darville, L., Kolluru, G. K., Rymond, C. C., Gerlach, B. D., Zheng, Z., et al. (2020) · Cell Metabolism · doi:10.1016/j.cmet.2020.01.001 · Cited by 525
Pessina, F., Giavazzi, F., Yin, Y., Gioia, U., Vitelli, V., Galbiati, A., Barozzi, S., Garre, M., Oldani, A., Flaus, A., et al. (2019) · Nature Cell Biology · doi:10.1038/s41556-019-0392-4 · Cited by 360
Metadata
- Authors
- Jao, C. Y., & Salic, A.
- Venue
- Proceedings of the National Academy of Sciences
- Published
- 2008-10-14
- Volume
- 105
- Issue
- 41
- Pages
- 15779-15784
- ISSN
- 0027-8424 (ISSN-L)
- 1091-6490
- OpenAlex
- W2136817553
- Cited by
- 693
- References
- 1
- FWCI
- 8.42
- Type
- journal article
- Language
- en
- Open access
- green
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