In situ detection of <i>JAK2</i>V617F within viable hematopoietic cells using gold nanoparticle technology

Sözer, Selçuk; Aptullahoglu, Erhan; Shivarov, Velizar; Yavuz, Akif Selim

doi:10.1111/ijlh.12991

Cited by 2 publications

(5 citation statements)

References 13 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…In order to ascertain whether c- MYC expression has an association with JAK2 V617F mutation, a previously established method for live cell analysis of JAK2 V617F mRNA and JAK2 wt transcripts with fluorochrome-conjugated gold nanoparticles [ 38 , 41 ] was applied to the MNCs of two Ph - MPN samples that had monoallelic JAK2 V617F. The JAK2 V617F mRNA- and JAK2wt mRNA-labeled gold nanoparticle-treated MNCs of MPN patients were sorted with a fluorescence-activated cell sorter for the JAK2 V617F and JAK2 wt transcripts, followed by analysis for relative c- MYC gene expression.…”

Section: Resultsmentioning

confidence: 99%

“…The transcript-labeled gold nanoparticle technology method was applied in this study for the mutant and wild-type allele-specific isolation of cells and distinct Ph - MPN MNC populations of JAK2 wt + and JAK2 V617F + were created [ 38 ]. This showed that the c -MYC , JAK2 , STAT5A , and STAT5B genes were overexpressed in JAK2 V617F + compared to JAK2 wt + .…”

Section: Discussionmentioning

confidence: 99%

“…Seven different probes were applied for the detection of JAK2 wild-type (wt) and JAK2 V617F transcripts in live cells as follows: two target-specific probes ( JAK2 wt-Cy5 and JAK2 V617F-Cy3) with a housekeeping control; beta-actin human-Cy3 and beta-actin human-Cy5; and three control probes (JAK2 Scramble-Cy3, Uptake-Cy3, and Scramble-Cy3+Uptake-Cy5) (EMD Millipore Company, Merck KGaA, Darmstadt, Germany), as explained elsewhere [ 38 ]. In brief, the MNCs were incubated with either the specific or control probes using 30000 cells at 200 µL/well in 96-well plates.…”

Section: Methodsmentioning

confidence: 99%

See 2 more Smart Citations

Circulating CD133+/–CD34– Have Increased c-MYC Expression in Myeloproliferative Neoplasms

Bıçak

Tokcan

Yavuz

et al. 2023

Tjh

View full text Add to dashboard Cite

Objective: Myeloproliferative neoplasms (MPNs) are hematopoietic stem cell (HSC)-originated diseases with clonal myeloproliferation. The constitutive activation of the JAK/STAT pathway is frequently detected in patients with Philadelphia chromosome-negative (Ph – ) MPNs with an acquired JAK2 V617F mutation. The c- MYC proto-oncogene is associated with malignant growth and cellular transformation, and JAK2 V617F was previously shown to induce constitutive expression of c- MYC . This study examines the expressional profile of c- MYC in Ph – MPNs with JAK2 V617F and highlights its hierarchical level of activation in circulating hematopoietic stem/progenitor cell (HSPC) subgroups. Materials and Methods: Mononuclear cells (MNCs) of Ph – MPNs were fluorochrome-labeled in situ with wild-type (wt) JAK2 or JAK2 V617F mRNA gold nanoparticle technology and sorted simultaneously. Isolated populations of JAK2 wt or JAK2 V617F were evaluated for their c- MYC expressions. The MNCs of 14 Ph – MPNs were further isolated for the study of HSPC subgroups regarding their CD34 and CD133 expressions, evaluated for the presence of JAK2 V617F, and compared to cord blood (CB) counterparts for the expression of c- MYC . Results: The mRNA-labeled gold nanoparticle-treated MNCs were determined to have the highest ratio of c- MYC relative fold-change expression in the biallelic JAK2 V617F compartment compared to JAK2 wt. The relative c- MYC expression in MNCs of MPNs was significantly increased compared to CB (p=0.01). The circulating HSPCs of CD133 +/– CD34 − MPNs had statistically significantly elevated c- MYC expression compared to CB. Conclusion: This is the first study of circulating CD133 +/– CD34 − cells in Ph – MPNs and it has revealed elevated c- MYC expression levels in HSCs/endothelial progenitor cells (HSCs/EPCs) and EPCs. Furthermore, the steady increase in the expression of c- MYC within MNCs carrying no mutations and monoallelic or biallelic JAK2 V617F transcripts was notable. The presence of JAK2 V617F with respect to c- MYC expression in the circulating HSCs/EPCs and EPCs of ...

show abstract

Section: Resultsmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

See 1 more Smart Citation

Circulating CD133+/–CD34– Have Increased c-MYC Expression in Myeloproliferative Neoplasms

Bıçak

Tokcan

Yavuz

et al. 2023

Tjh

View full text Add to dashboard Cite

show abstract

“…Scrambled NanoFlares with no target in the organism showed no fluorescence after incubation, while positive control NanoFlares targeting poly A RNA fluoresced throughout the organism, as expected. In another successful use of NanoFlares, Sozer et al have recently used NanoFlares to detect single nucleotide mutations in Janus kinase 2 mRNA …”

Section: Hybridization‐based Probesmentioning

confidence: 99%

“…In another successful use of Nano-Flares, Sozer et al have recently used NanoFlares to detect single nucleotide mutations in Janus kinase 2 mRNA. [173] Slight variations in the original NanoFlare design have led to NanoFlares with additional capabilities. For example, multiplexed NanoFlares were developed in 2012 to detect multiple mRNA transcripts with a single construct.…”

Section: Gold Nanoparticle Basedmentioning

confidence: 99%

Nucleic‐Acid Structures as Intracellular Probes for Live Cells

2019

View full text Add to dashboard Cite

The chemical composition of cells at the molecular level determines their growth, differentiation, structure, and function. Probing this composition is powerful because it provides invaluable insight into chemical processes inside cells and in certain cases allows disease diagnosis based on molecular profiles. However, many techniques analyze fixed cells or lysates of bulk populations, in which information about dynamics and cellular heterogeneity is lost. Recently, nucleic‐acid‐based probes have emerged as a promising platform for the detection of a wide variety of intracellular analytes in live cells with single‐cell resolution. Recent advances in this field are described and common strategies for probe design, types of targets that can be identified, current limitations, and future directions are discussed.

show abstract

In situ detection of JAK2V617F within viable hematopoietic cells using gold nanoparticle technology

Cited by 2 publications

References 13 publications

Circulating CD133+/–CD34– Have Increased c-MYC Expression in Myeloproliferative Neoplasms

Circulating CD133+/–CD34– Have Increased c-MYC Expression in Myeloproliferative Neoplasms

Nucleic‐Acid Structures as Intracellular Probes for Live Cells

Contact Info

Product

Resources

About