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| Item Type: | Article |
|---|---|
| Title: | Comparing agent‐based delivery of DNA and PNA forced intercalation (FIT) probes for multicolor mRNA imaging |
| Creators Name: | Chamiolo, J. and Fang, G.M. and Hövelmann, F. and Friedrich, D. and Knoll, A. and Loewer, A. and Seitz, O. |
| Abstract: | Fluorogenic oligonucleotide probes allow mRNA imaging in living cells. A key challenge is the cellular delivery of probes. Most delivery agents, such as cell-penetrating peptides (CPPs) and pore-forming proteins, require interactions with the membrane. Charges play an important role. To explore the influence of charge on fluorogenic properties and delivery efficiency, we compared peptide nucleic acid (PNA)- with DNA-based forced intercalation (FIT) probes. Perhaps counterintuitively, fluorescence signaling by charged DNA FIT probes proved tolerant to CPP conjugation, whereas CPP–FIT PNA conjugates were affected. Live-cell imaging was performed with a genetically engineered HEK293 cell line to allow the inducible expression of a specific mRNA target. Blob-like features and high background were recurring nuisances of the tested CPP and lipid conjugates. By contrast, delivery by streptolysin-O provided high enhancements of the fluorescence of the FIT probe upon target induction. Notably, DNA-based FIT probes were brighter and more responsive than PNA-based FIT probes. Optimized conditions enabled live-cell multicolor imaging of three different mRNA target sequences. |
| Keywords: | Bioconjugation, Fluorescence, Imaging Agents, Oligonucleotides, RNA |
| Source: | ChemBioChem |
| ISSN: | 1439-4227 |
| Publisher: | Wiley-VCH |
| Volume: | 20 |
| Number: | 4 |
| Page Range: | 595-604 |
| Date: | 15 February 2019 |
| Official Publication: | https://doi.org/10.1002/cbic.201800526 |
| PubMed: | View item in PubMed |
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