Journal Article

PUBDB-2024-00347

http://join2-wiki.gsi.de/foswiki/pub/Main/Artwork/join2_logo100x88.png Concatemeric Broccoli reduces mRNA stability and induces aggregates

Rink, M. R. ; Baptista, M. A. P. ; Flomm, F. J.Extern* ; Hennig, T. ; Whisnant, A. W. ; Wolf, N. ; Seibel, J. ; Dölken, L. ; Bosse, J. B. (Corresponding author)Extern*CSSB*

2021
PLOS San Francisco, California, US

This record in other databases:        

Please use a persistent id in citations: doi:10.1371/journal.pone.0244166  doi:10.3204/PUBDB-2024-00347

Abstract: Fluorogenic aptamers are an alternative to established methodology for real-time imaging of RNA transport and dynamics. We developed Broccoli-aptamer concatemers ranging from 4 to 128 substrate-binding site repeats and characterized their behavior fused to an mCherry-coding mRNA in transient transfection, stable expression, and in recombinant cytomegalovirus infection. Concatemerization of substrate-binding sites increased Broccoli fluorescence up to a concatemer length of 16 copies, upon which fluorescence did not increase and mCherry signals declined. This was due to the combined effects of RNA aptamer aggregation and reduced RNA stability. Unfortunately, both cellular and cytomegalovirus genomes were unable to maintain and express high Broccoli concatemer copy numbers, possibly due to recombination events. Interestingly, negative effects of Broccoli concatemers could be partially rescued by introducing linker sequences in between Broccoli repeats warranting further studies. Finally, we show that even though substrate-bound Broccoli is easily photobleached, it can still be utilized in live-cell imaging by adapting a time-lapse imaging protocol.

---

Contributing Institute(s):

1. CSSB-MHH-JB (CSSB-MHH-JB)

Research Program(s):

1. 899 - ohne Topic (POF4-899) (POF4-899)
2. DFG project 390874280 - EXC 2155: RESIST - Resolving Infection Susceptibility (390874280) (390874280)

Experiment(s):

1. No specific instrument

---

Database coverage:
;

---