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Comparing agent‐based delivery of DNA and PNA forced intercalation (FIT) probes for multicolor mRNA imaging

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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 (Germany)
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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