5-Ethynyl-UTP (5-EUTP) can be incorporated into RNA using standard nucleotide mixtures and T7 RNA polymerase in vitro transcription. For a partial modified RNA we recommend to replace the natural base by only 1-10%. Modified mRNA is ready for labeling with marker azides (e.g. fluorescent dyes for tracking and imaging or cross-linking to other biomolecules) by click chemistry.

LITERATURE

Copper-Catalyzed Azide–Alkyne Click Chemistry for Bioconjugation, S. Presolski et al., 2011, Current Protocols of Chemical Biology, Vol. 3, p. 153-162.

https://doi.org/10.1002/9780470559277.ch110148

Chemoenzymatic Preparation of Functional Click-Labeled Messenger RNA, S. Croce et al., 2020, ChemBioChem, Vol. 21, p. 1641-1646.

https://doi.org/10.1002/cbic.201900718

ALS-linked FUS mutants affect the localization of U7 snRNP and replication-dependent histone gene expression in human cells, A. Gadgil et al., 2021, Scientific Reports, Vol. 11(1), p. 11868.

https://doi.org/10.1038/s41598-021-91453-3