Metabolic RNA labeling in non-engineered cells following spontaneous uptake of fluorescent nucleoside phosphate analogues
Artikel i vetenskaplig tidskrift, 2024

RNA and its building blocks play central roles in biology and have become increasingly important as therapeutic agents and targets. Hence, probing and understanding their dynamics in cells is important. Fluorescence microscopy offers live-cell spatiotemporal monitoring but requires labels. We present two fluorescent adenine analogue nucleoside phosphates which show spontaneous uptake and accumulation in cultured human cells, likely via nucleoside transporters, and show their potential utilization as cellular RNA labels. Upon uptake, one nucleotide analogue, 2CNqAXP, localizes to the cytosol and the nucleus. We show that it could then be incorporated into de novo synthesized cellular RNA, i.e. it was possible to achieve metabolic fluorescence RNA labeling without using genetic engineering to enhance incorporation, uptake-promoting strategies, or post-labeling through bio-orthogonal chemistries. By contrast, another nucleotide analogue, pAXP, only accumulated outside of the nucleus and was rapidly excreted. Consequently, this analogue did not incorporate into RNA. This difference in subcellular accumulation and retention results from a minor change in nucleobase chemical structure. This demonstrates the importance of careful design of nucleoside-based drugs, e.g. antivirals to direct their subcellular localization, and shows the potential of fine-tuning fluorescent base analogue structures to enhance the understanding of the function of such drugs.

Författare

Pauline Pfeiffer

Chalmers, Kemi och kemiteknik, Kemi och biokemi

Jesper Nilsson

Chalmers, Kemi och kemiteknik, Kemi och biokemi

Audrey Gallud

Chalmers, Life sciences, Kemisk biologi

AstraZeneca AB

Tom Baladi

Chalmers, Kemi och kemiteknik, Kemi och biokemi

AstraZeneca AB

Hoang Ngoan Le

Chalmers, Kemi och kemiteknik, Kemi och biokemi

AstraZeneca AB

Mattias Bood

AstraZeneca AB

Göteborgs universitet

Malin Lemurell

AstraZeneca AB

Anders Dahlén

AstraZeneca AB

Morten Grötli

Göteborgs universitet

Elin Esbjörner Winters

Chalmers, Life sciences, Kemisk biologi

Marcus Wilhelmsson

Chalmers, Kemi och kemiteknik, Kemi och biokemi

Nucleic Acids Research

0305-1048 (ISSN) 1362-4962 (eISSN)

Vol. 52 17 10102-10118

Funktionell leverans av nukleotid-baserade läkemedel

Stiftelsen för Strategisk forskning (SSF) (IRC15-0065), 2017-03-01 -- 2024-12-31.

Endogennaturlik fluorescensmärkning villkor av RNA och dess användning i cellavbildning

Vetenskapsrådet (VR) (2021-04409), 2021-12-01 -- 2025-11-30.

Styrkeområden

Nanovetenskap och nanoteknik

Ämneskategorier

Biokemi och molekylärbiologi

DOI

10.1093/nar/gkae722

PubMed

39162218

Mer information

Senast uppdaterat

2024-12-06