Antisense Oligonucleotides Used to Identify Telomeric G-Quadruplexes in Metaphase Chromosomes and Fixed Cells by Fluorescence Lifetime Imaging Microscopy of o-BMVC Foci

Tseng, Ting-Yuan; Liu, Shin-Ya; Wang, Chiung-Lin; Chang, Ta‐Chau

doi:10.3390/molecules25184083

Cited by 8 publications

(5 citation statements)

References 65 publications

(77 reference statements)

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“…Based on the distinctive fluorescence lifetime response of G-Quadruplex, the content of DNA G-Quadruplex in lung, mammary and hepatic cancer cells is four times than that in normal cells, which can be exploited for cancer diagnosis [ 104 ]. Similarly, another G-Quadruplex fluorescent probe called o-BMVC foci, has also been used to identify G-Quadruplex motifs in combination with FLIM [ 105 ]. In addition to G-Quadruplex, the acetylation of histone lysine residues of acetyltransferase is another target for anti-cancer drugs that can be monitored by using a P300/CBP associated factor (PCAF) biosensor (HATS) in combination with FLIM (Fig.…”

Section: Flim Combined With Fluorophore Dye Probesmentioning

confidence: 99%

FLIM as a Promising Tool for Cancer Diagnosis and Treatment Monitoring

et al. 2021

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Fluorescence lifetime imaging microscopy (FLIM) has been rapidly developed over the past 30 years and widely applied in biomedical engineering. Recent progress in fluorophore-dyed probe design has widened the application prospects of fluorescence. Because fluorescence lifetime is sensitive to microenvironments and molecule alterations, FLIM is promising for the detection of pathological conditions. Current cancer-related FLIM applications can be divided into three main categories: (i) FLIM with autofluorescence molecules in or out of a cell, especially with reduced form of nicotinamide adenine dinucleotide, and flavin adenine dinucleotide for cellular metabolism research; (ii) FLIM with Förster resonance energy transfer for monitoring protein interactions; and (iii) FLIM with fluorophore-dyed probes for specific aberration detection. Advancements in nanomaterial production and efficient calculation systems, as well as novel cancer biomarker discoveries, have promoted FLIM optimization, offering more opportunities for medical research and applications to cancer diagnosis and treatment monitoring. This review summarizes cutting-edge researches from 2015 to 2020 on cancer-related FLIM applications and the potential of FLIM for future cancer diagnosis methods and anti-cancer therapy development. We also highlight current challenges and provide perspectives for further investigation.

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Section: Flim Combined With Fluorophore Dye Probesmentioning

confidence: 99%

FLIM as a Promising Tool for Cancer Diagnosis and Treatment Monitoring

et al. 2021

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“…3). FLIM has been applied to quantitatively measure the number of G4 foci in metaphase chromosomes from number of cell lines showing that G4 structures are formed by human telomeric TTAGGG repeats on chromosomes (Tseng et al 2020). FLIM-FRET in vivo has shown that the concentration of polyvalent cations and ATP levels affect chromatin conformation and compaction.…”

Section: Introductionmentioning

confidence: 99%

Contribution of advanced fluorescence nano microscopy towards revealing mitotic chromosome structure

et al. 2021

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The organization of chromatin into higher-order structures and its condensation process represent one of the key challenges in structural biology. This is important for elucidating several disease states. To address this long-standing problem, development of advanced imaging methods has played an essential role in providing understanding into mitotic chromosome structure and compaction. Amongst these are two fast evolving fluorescence imaging technologies, specifically fluorescence lifetime imaging (FLIM) and super-resolution microscopy (SRM). FLIM in particular has been lacking in the application of chromosome research while SRM has been successfully applied although not widely. Both these techniques are capable of providing fluorescence imaging with nanometer information. SRM or “nanoscopy” is capable of generating images of DNA with less than 50 nm resolution while FLIM when coupled with energy transfer may provide less than 20 nm information. Here, we discuss the advantages and limitations of both methods followed by their contribution to mitotic chromosome studies. Furthermore, we highlight the future prospects of how advancements in new technologies can contribute in the field of chromosome science.

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“…The use of an antisense sequence was previously applied to unfold human telomeric G4 structures in vitro [ 27 , 28 ] and in vivo [ 29 ]. In the presence of antisense DNA, some of the unfolded species that formed as a result of thermal motion can be trapped as a stable duplex.…”

Section: Resultsmentioning

confidence: 99%

Folding and Unfolding of Exogenous G-Rich Oligonucleotides in Live Cells by Fluorescence Lifetime Imaging Microscopy of o-BMVC Fluorescent Probe

et al. 2021

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Guanine-rich oligonucleotides (GROs) can self-associate to form G-quadruplex (G4) structures that have been extensively studied in vitro. To translate the G4 study from in vitro to in live cells, here fluorescence lifetime imaging microscopy (FLIM) of an o-BMVC fluorescent probe is applied to detect G4 structures and to study G4 dynamics in CL1-0 live cells. FLIM images of exogenous GROs show that the exogenous parallel G4 structures that are characterized by the o-BMVC decay times (≥2.4 ns) are detected in the lysosomes of live cells in large quantities, but the exogenous nonparallel G4 structures are hardly detected in the cytoplasm of live cells. In addition, similar results are also observed for the incubation of their single-stranded GROs. In the study of G4 formation by ssHT23 and hairpin WT22, the analyzed binary image can be used to detect very small increases in the number of o-BMVC foci (decay time ≥ 2.4 ns) in the cytoplasm of live cells. However, exogenous ssCMA can form parallel G4 structures that are able to be detected in the lysosomes of live CL1-0 cells in large quantities. Moreover, the photon counts of the o-BMVC signals (decay time ≥ 2.4 ns) that are measured in the FLIM images are used to reveal the transition of the G4 formation of ssCMA and to estimate the unfolding rate of CMA G4s with the addition of anti-CMA into live cells for the first time. Hence, FLIM images of o-BMVC fluorescence hold great promise for the study of G4 dynamics in live cells.

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Antisense Oligonucleotides Used to Identify Telomeric G-Quadruplexes in Metaphase Chromosomes and Fixed Cells by Fluorescence Lifetime Imaging Microscopy of o-BMVC Foci

Cited by 8 publications

References 65 publications

FLIM as a Promising Tool for Cancer Diagnosis and Treatment Monitoring

FLIM as a Promising Tool for Cancer Diagnosis and Treatment Monitoring

Contribution of advanced fluorescence nano microscopy towards revealing mitotic chromosome structure

Folding and Unfolding of Exogenous G-Rich Oligonucleotides in Live Cells by Fluorescence Lifetime Imaging Microscopy of o-BMVC Fluorescent Probe

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