Optical Antisense Tumor Targeting in Vivo with an Improved Fluorescent DNA Duplex Probe

Liu, Xinrong; Cheng, Dengfeng; Nakamura, Kayoko; Wang, Yi; Dou, Shuping; Liu, Guozheng; Rusckowski, Mary; Hnatowich, Donald J.

doi:10.1021/bc9000933

Cited by 11 publications

(15 citation statements)

References 22 publications

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“…Thiazole Orange is a commercially available, high quantum yield fluorophore that emits strong fluorescence upon binding to nucleic acids, resulting in highly sensitive nucleic acid sensing and DNA imaging . This fluorophore has been conjugated to peptides, DNA, RNA, and other oligonucleotide variants, and its high cell permeability and intracellular stability means that these tagged macromolecules can be readily imaged in vivo. , Because the fluorescence of Thiazole Orange is highly sensitive to the presence of exogenous contaminants, it can be used for the detection of a broad variety of analytes, including tetracycline, an antibiotic; cations; , melamine, a highly toxic compound used in the production of plastics; and other analytes of interest. , …”

Section: Common Fluorophoresmentioning

confidence: 99%

Supramolecular Luminescent Sensors

2018

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There is great need for stand-alone luminescence-based chemosensors that exemplify selectivity, sensitivity, and applicability and that overcome the challenges that arise from complex, real-world media. Discussed herein are recent developments toward these goals in the field of supramolecular luminescent chemosensors, including macrocycles, polymers, and nanomaterials. Specific focus is placed on the development of new macrocycle hosts since 2010, coupled with considerations of the underlying principles of supramolecular chemistry as well as analytes of interest and common luminophores. State-of-the-art developments in the fields of polymer and nanomaterial sensors are also examined, and some remaining unsolved challenges in the area of chemosensors are discussed. CONTENTS 4.4. Explosives 4.5. Pharmaceutical Agents 4.6. Biologically Relevant 4.6.1. Adenosine Triphosphate (ATP)

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Section: Common Fluorophoresmentioning

confidence: 99%

Supramolecular Luminescent Sensors

2018

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“…Based on the experience of this laboratory with numerous oligomer backbone chemistries [9,19,20,29], and the work of others [30,31], the three oligomer backbone chemistries selected for these studies were PS-DNA, PNA and MORF.…”

Section: Discussionmentioning

confidence: 99%

“…Since the phosphodiester DNA is unstable to nucleases [17], and since the pharmacokinetics and binding properties of oligomers can depend on their structure [18] three different oligomer types were studied as alternatives to the native phosphodiester DNA: PNA; phosphorothioate DNA (PS-DNA) and MORF. Each oligomer type has previously been radiolabeled in this laboratory with 99m Tc for various applications [9,10,19,20]. These oligomers differ in the linkages between the bases and in charge, but each is stable to nucleases and each maintains the proper structure for complementary base pairing and stable hybridization.…”

Section: Introductionmentioning

confidence: 99%

99mTc-MORF oligomers specific for bacterial ribosomal RNA as potential specific infection imaging agents

Chen

Wang

Cheng

et al. 2013

Bioorganic & Medicinal Chemistry

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Purpose Radiolabeled oligomers complementary to the 16S rRNA in bacteria were investigated as bacterial infection imaging agents. Methods and Results Identical sequences with backbones phosphorodiamidate morpholino (MORF), peptide nucleic acid (PNA), and phosphorothioate DNA (PS-DNA) were 99mTc-labeled and evaluated for binding to bacterial RNA. MORF binding to RNA from E. coli strains SM101 and K12 was 4-fold and 150-fold higher compared to PNA, and PS-DNA, respectively. Subsequently MORF oligomer in fluorescence in situ hybridization showed a stronger signal with study MORF compared to control in fixed preparations of two E. coli strains and K. pneumoniae. Flow cytometry analysis showed study MORF accumulation to be 8- and 80-fold higher compared to the control in live K. pneumoniae and S. aureus, respectively. Further, fluorescence microscopy showed increased accumulation of study MORF over control in live E. coli and K. pneumonia. Binding of 99mTc-study MORF to RNA from E. coli SM101 and K12 was 30.4 pmoles and 117.8 pmoles, respectively, per 1010 cells. Mice with K. pneumoniae live or heat-killed (sterile inflammation) in one thigh at 90 min for both 99mTc-study MORF and control showed higher accumulation in target thighs than in blood and all organs other than kidney and small intestines. Whereas accumulation of 99mTc-study MORF was significantly higher (p=0.009) than that of the control in the thigh with sterile inflammation. Conclusion A 99mTc-MORF oligomer complimentary to the bacterial 16S rRNA demonstrated binding to bacterial RNA in vitro with specific accumulation into live bacteria. Radiolabeled MORF oligomers antisense to the bacterial rRNA may be useful to image bacterial infection.

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“…They showed that this probe could image a complementary biotinylated 25-mer MORF oligomer immobilized on streptavidin polystyrene microspheres that were intramuscularly implanted into a mouse [ 14 ]. The same group also utilized a probe consisting of a 25-mer phosphorothioate DNA bearing Cy5.5 and a 10-mer complementary ODN with the BHQ3 quencher to image the KB-G2 tumor in mice which overexpresses the multi-drug-resistant mdr1 mRNA [ 15 ]. In another approach, Mirkin and coworkers developed “nanoflares” in which antisense ODNs to a target mRNA are conjugated to a gold nanoparticle and then hybridized to a shorter strand of complementary DNA bearing Cy5 which is quenched by the gold nanoparticle.…”

Section: Introductionmentioning

confidence: 99%

Imaging mRNA Expression in Live Cells via PNA·DNA Strand Displacement-Activated Probes

Wang

Zhang

Wooley

et al. 2012

Journal of Nucleic Acids

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Probes for monitoring mRNA expression in vivo are of great interest for the study of biological and biomedical problems, but progress has been hampered by poor signal to noise and effective means for delivering the probes into live cells. Herein we report a PNA·DNA strand displacement-activated fluorescent probe that can image the expression of iNOS (inducible nitric oxide synthase) mRNA, a marker of inflammation. The probe consists of a fluorescein labeled antisense PNA annealed to a shorter DABCYLplus-labeled DNA which quenches the fluorescence, but when the quencher strand is displaced by the target mRNA the fluorescence is restored. DNA was used for the quencher strand to facilitate electrostatic binding of the otherwise netural PNA strand to a cationic shell crosslinked knedel-like (cSCK) nanoparticle which can deliver the PNA·DNA duplex probe into cells with less toxicity and greater efficiency than other transfection agents. RAW 264.7 mouse macrophage cells transfected with the iNOS PNA·DNA probe via the cSCK showed a 16 to 54-fold increase in average fluorescence per cell upon iNOS stimulation. The increase was 4 to 7-fold higher than that for a non-complementary probe, thereby validating the ability of a PNA·DNA strand displacement-activated probe to image mRNA expression in vivo.

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Optical Antisense Tumor Targeting in Vivo with an Improved Fluorescent DNA Duplex Probe

Cited by 11 publications

References 22 publications

Supramolecular Luminescent Sensors

Supramolecular Luminescent Sensors

99mTc-MORF oligomers specific for bacterial ribosomal RNA as potential specific infection imaging agents

Imaging mRNA Expression in Live Cells via PNA·DNA Strand Displacement-Activated Probes

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