Photoinduzierter reduktiver Elektronentransfer in LNA:DNA‐Hybriden: Kompromiss aus Konformation und Basenstapelung

Bei der Analyse der RNA-Dynamik in lebenden Zellen werden üblicherweise transgene Methoden eingesetzt. Diese erfordern modifizierte RNAs und Zellen. Hingegen ermçglichen Hybridisierungs-Fluoreszenzsonden Untersuchungen an Wildtyp-Zellen. Wir haben Nuklease-resistente FIT("DNA-forced intercalation")-Sonden entwickelt, in denen hohe Anstiege der Fluoreszenz durch Hybridisierung mit einer großen Helligkeit einhergehen, sodass einzelne Ribonukleotidpartikel (RNP) verfolgt werden kçnnen. Hierbei dient ein einzelner Thiazolorange(TO)-Interkalationsfarbstoff als Nukleobasensurrogat während eine benachbarte LNA("locked nucleic acid")-Modifikation die Umgebung konformativ fixiert. Dies schließt Fluoreszenzabklingkanäle und erhçht so die Helligkeit von TO in Sonden-Ziel-Komplexen. Zwei FIT-Sonden reichen aus, um oskar-RNPs in lebenden Wildtyp-Oozyten von Drosophila melanogaster zu verfolgen.

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Helligkeit durch lokale Rigidifizierung – LNA‐verstärkte FIT‐Sonden zur bildgebenden Darstellung von Ribonukleotidpartikeln in vivo

Hövelmann

Gáspár

Loibl

et al. 2014

Angewandte Chemie

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Reductive Charge Transfer through an RNA Aptamer

Frommer

Müller

2020

Angewandte Chemie

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The transfer of charges through double helical DNA is a very well investigated bioelectric phenomenon. RNA, on the contrary, has been less studied in this regard. The few available data report on charge transfer through RNA duplex structures mainly composed of homonucleotide sequences. In the light of the RNA world scenarios, it is an interesting question, if charge transfer can be coupled with RNA function. Functional RNAs however, contain versatile structural motifs. Therefore, electron transport also through non-Watson-Crick base-paired regions might be required. We here demonstrate distance-dependent reductive charge transfer through RNA duplexes and through the non-Watson-Crick base-paired region of an RNA aptamer.

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Brightness through Local Constraint—LNA‐Enhanced FIT Hybridization Probes for In Vivo Ribonucleotide Particle Tracking

et al. 2014

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Imaging the dynamics of RNA in living cells is usually performed by means of transgenic approaches that require modification of RNA targets and cells. Fluorogenic hybridization probes would also allow the analysis of wild-type organisms. We developed nuclease-resistant DNA forced intercalation (FIT) probes that combine the high enhancement of fluorescence upon hybridization with the high brightness required to allow tracking of individual ribonucleotide particles (RNPs). In our design, a single thiazole orange (TO) intercalator dye is linked as a nucleobase surrogate and an adjacent locked nucleic acid (LNA) unit serves to introduce a local constraint. This closes fluorescence decay channels and thereby increases the brightness of the probe-target duplexes. As few as two probes were sufficient to enable the tracking of oskar mRNPs in wild-type living Drosophila melanogaster oocytes.

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Photoinduzierter reduktiver Elektronentransfer in LNA:DNA‐Hybriden: Kompromiss aus Konformation und Basenstapelung

Cited by 6 publications

References 48 publications

Helligkeit durch lokale Rigidifizierung – LNA‐verstärkte FIT‐Sonden zur bildgebenden Darstellung von Ribonukleotidpartikeln in vivo

Helligkeit durch lokale Rigidifizierung – LNA‐verstärkte FIT‐Sonden zur bildgebenden Darstellung von Ribonukleotidpartikeln in vivo

Reductive Charge Transfer through an RNA Aptamer

Brightness through Local Constraint—LNA‐Enhanced FIT Hybridization Probes for In Vivo Ribonucleotide Particle Tracking

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