Fluorescence Imaging of Influenza H1N1 mRNA in Living Infected Cells Using Single‐Chromophore FIT‐PNA

Kummer, Susann; Knoll, Andrea; Socher, Elke; Bethge, Lucas; Herrmann, Andreas; Seitz, Oliver

doi:10.1002/anie.201005902

Cited by 113 publications

(84 citation statements)

References 35 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The TO dye is able to behave as universal base (41). Moreover, the fluorescence emission of the TO-PNA monomer in the context of a PNA ON is enhanced upon binding to a complementary DNA or RNA strand in vitro (41–43) and in cells (43). We carried out a scan of K-PNA19mer-K3 anti-miR by replacement of individual PNA units at different sequence positions by the TO-PNA monomer and by measurement of the fluorescence differences when the PNA is single-stranded (ss-PNA) or in a duplex with a complementary miR-122 RNA (PNA–RNA duplex) at 25 or 37°C (Supplementary Figure S1B and C).…”

Section: Resultsmentioning

confidence: 99%

Chemical structure requirements and cellular targeting of microRNA-122 by peptide nucleic acids anti-miRs

Torres

Fabani

Vigorito

et al. 2011

Nucleic Acids Research

Self Cite

104

View full text Add to dashboard Cite

Anti-miRs are oligonucleotide inhibitors complementary to miRNAs that have been used extensively as tools to gain understanding of specific miRNA functions and as potential therapeutics. We showed previously that peptide nucleic acid (PNA) anti-miRs containing a few attached Lys residues were potent miRNA inhibitors. Using miR-122 as an example, we report here the PNA sequence and attached amino acid requirements for efficient miRNA targeting and show that anti-miR activity is enhanced substantially by the presence of a terminal-free thiol group, such as a Cys residue, primarily due to better cellular uptake. We show that anti-miR activity of a Cys-containing PNA is achieved by cell uptake through both clathrin-dependent and independent routes. With the aid of two PNA analogues having intrinsic fluorescence, thiazole orange (TO)-PNA and [bis-o-(aminoethoxy)phenyl]pyrrolocytosine (BoPhpC)-PNA, we explored the subcellular localization of PNA anti-miRs and our data suggest that anti-miR targeting of miR-122 may take place in or associated with endosomal compartments. Our findings are valuable for further design of PNAs and other oligonucleotides as potent anti-miR agents.

show abstract

Section: Resultsmentioning

confidence: 99%

Chemical structure requirements and cellular targeting of microRNA-122 by peptide nucleic acids anti-miRs

Torres

Fabani

Vigorito

et al. 2011

Nucleic Acids Research

Self Cite

104

View full text Add to dashboard Cite

show abstract

“…[112][113][114] The authors successfully applied the probes to the detection of influenza H1N1 mRNA in living infected cells, which represents the first example of an artificial nucleotidic probe that can be used for imaging of mRNA in living cells. 115 The probes achieved an 11-fold increase in TO emission upon hybridization with the complementary RNA target.…”

Section: Fluorescence Imaging With Nucleic Acid Probesmentioning

confidence: 99%

“…116,117 A commonly used nucleic acid templated reaction is Staudinger ligation, a fast reaction between azide and phosphine. 115,118 The remarkable advantage of this type of probe is that the reaction only occurs when the two reactive groups are anchored with close proximity, and thus fluorescence enhancement can only be observed in the presence of the target nucleic acids. Moreover, these probes also share other benefits such as fast reaction kinetics and improved signal amplification.…”

Section: Fluorescence Imaging With Nucleic Acid Probesmentioning

confidence: 99%

Nucleic Acid Fluorescent Probes for Biological Sensing

Xiao

Zhang

et al. 2012

Appl Spectrosc

View full text Add to dashboard Cite

Nucleic acid fluorescent probes are playing increasingly important roles in biological sensing in recent years. In addition to the conventional functions of single-stranded DNA/RNA to hybridize with their complementary strands, affinity nucleic acids (aptamers) with specific target binding properties have also been developed, which has greatly broadened the application of nucleic acid fluorescent probes to the detection of a large variety of analytes, including small molecules, proteins, ions, and even whole cells. Another chemical property of nucleic acids is to act as substrates for various nucleic acid enzymes. This property can be utilized not only to detect those enzymes and screen their inhibitors, but also employed to develop effective signal amplification systems, which implies extensive applications. This review mainly covers the biosensing methods based on the above three types of nucleic acid fluorescent probes. The most widely used intensity-based biosensing assays are covered first, including nucleic acid probe-based signal amplification methods. Then fluorescence lifetime, fluorescence anisotropy, and fluorescence correlation spectroscopy assays are introduced, respectively. As a rapidly developing field, fluorescence imaging approaches are also briefly summarized.

show abstract

“…11a). Kummer et al 143 used a PNA-FIT probe to image Influenza H1N1 mRNA, and the fluorescence of the probe increased 11-fold in the absence of the target RNAs, while that of the control MB probe targeting the same RNA only increased 6-fold. The high fluorescence increase is attributed to the properties of TO in the probe as well as the PNA structures, which have higher affinity and better biostability than common DNA structures.…”

Section: Probes Imaging Endogenous Rnasmentioning

confidence: 99%

Recent advances in high-performance fluorescent and bioluminescent RNA imaging probes

et al. 2017

View full text Add to dashboard Cite

RNA plays an important role in life processes. Imaging of messenger RNAs (mRNAs) and micro-RNAs (miRNAs) not only allows us to learn the formation and transcription of mRNAs and the biogenesis of miRNAs involved in various life processes, but also helps in detecting cancer. High-performance RNA imaging probes greatly expand our view of life processes and enhance the cancer detection accuracy. In this review, we summarize the state-of-the-art high-performance RNA imaging probes, including exogenous probes that can image RNA sequences with special modification and endogeneous probes that can directly image endogenous RNAs without special treatment. For each probe, we review its structure and imaging principle in detail. Finally, we summarize the application of mRNA and miRNA imaging probes in studying life processes as well as in detecting cancer. By correlating the structures and principles of various probes with their practical uses, we compare different RNA imaging probes and offer guidance for better utilization of the current imaging probes and the future design of higher-performance RNA imaging probes.

show abstract

Fluorescence Imaging of Influenza H1N1 mRNA in Living Infected Cells Using Single‐Chromophore FIT‐PNA

Cited by 113 publications

References 35 publications

Chemical structure requirements and cellular targeting of microRNA-122 by peptide nucleic acids anti-miRs

Chemical structure requirements and cellular targeting of microRNA-122 by peptide nucleic acids anti-miRs

Nucleic Acid Fluorescent Probes for Biological Sensing

Recent advances in high-performance fluorescent and bioluminescent RNA imaging probes

Contact Info

Product

Resources

About