Young-Kug Choo's research while affiliated with Wonkwang University and other places

What is this page?


This page lists the scientific contributions of an author, who either does not have a ResearchGate profile, or has not yet added these contributions to their profile.

It was automatically created by ResearchGate to create a record of this author's body of work. We create such pages to advance our goal of creating and maintaining the most comprehensive scientific repository possible. In doing so, we process publicly available (personal) data relating to the author as a member of the scientific community.

If you're a ResearchGate member, you can follow this page to keep up with this author's work.

If you are this author, and you don't want us to display this page anymore, please let us know.

Publications (107)


Quercetin Induces Mitochondrial Apoptosis and Downregulates Ganglioside GD3 Expression in Melanoma Cells
  • Article
  • Full-text available

May 2024

·

13 Reads

International Journal of Molecular Sciences

Sang Young Seo

·

Won Seok Ju

·

Kyongtae Kim

·

[...]

·

Young-Kug Choo

Malignant melanoma represents a form of skin cancer characterized by a bleak prognosis and heightened resistance to traditional therapies. Quercetin has demonstrated notable anti-carcinogenic, anti-inflammatory, anti-oxidant, and pharmacological effects across various cancer types. However, the intricate relationship between quercetin’s anti-cancer properties and ganglioside expression in melanoma remains incompletely understood. In this study, quercetin manifests specific anti-proliferative, anti-migratory, and cell-cycle arrest effects, inducing mitochondrial dysfunction and apoptosis in two melanoma cancer cell lines. This positions quercetin as a promising candidate for treating malignant melanoma. Moreover, our investigation indicates that quercetin significantly reduces the expression levels of ganglioside GD3 and its synthetic enzyme. Notably, this reduction is achieved through the inhibition of the FAK/paxillin/Akt signaling pathway, which plays a crucial role in cancer development. Taken together, our findings suggest that quercetin may be a potent anti-cancer drug candidate for the treatment of malignant melanoma.

Download
Share

Figure 4. Effect of treatment with the ganglioside GD3 inhibitor NEU3 on TNF-α-treated hNECs. (A) ST8SIA1 gene expression in TNF-α-treated hNECs with or without NEU3 treatment. (B) Highperformance thin-layer chromatography analysis of ganglioside expression in TNF-α-treated hNECs with or without NEU3 treatment. M1 and M2 are ganglioside standard markers. (C) mRNA expression levels of inflammation-related genes in TNF-α-treated hNECs with or without NEU3 treatment. (D) NF-κB/P65 (cytoplasmic and nuclear) were analyzed by Western blotting. (E) mRNA and (F) protein expression of epithelial-to-mesenchymal transition-related genes in TNF-α-treated hNECs with or without NEU3 treatment. mRNA and protein expression levels were normalized to that of the housekeeping gene β-actin. (G) Wound healing assay. hNECs were placed on 6-well plate and were treated with or without NEU3 (50 ng/mL) for 1 h and wounded by a blue pipette tip. And then treated with 20 ng/mL of TNF-α for 24 h. Scale bar 100 µm. Expression levels are presented as the mean ± SD from three independent experiments and analyzed using Student's t-test. *p < 0.05,
Clinical characteristics of patients included in this study.
Ganglioside GD3 Regulates Inflammation and Epithelial-to-Mesenchymal Transition in Human Nasal Epithelial Cells

April 2024

·

18 Reads

International Journal of Molecular Sciences

Chronic sinusitis with nasal polyps (CRSwNP) is one of the most common chronic inflammatory diseases, and involves tissue remodeling. One of the key mechanisms of tissue remodeling is the epithelial-mesenchymal transition (EMT), which also represents one of the pathophysiological processes of CRS observed in CRSwNP tissues. To date, many transcription factors and forms of extracellular stimulation have been found to regulate the EMT process. However, it is not known whether gangliosides, which are the central molecules of plasma membranes, involved in regulating signal transmission pathways, are involved in the EMT process. Therefore, we aimed to determine the role of gangliosides in the EMT process. First, we confirmed that N-cadherin, which is a known mesenchymal marker, and ganglioside GD3 were specifically expressed in CRSwNP_NP tissues. Subsequently, we investigated whether the administration of TNF-α to human nasal epithelial cells (hNECs) resulted in the upregulation of ganglioside GD3 and its synthesizing enzyme, ST8 alpha-N-acetyl-neuraminide alpha-2,8-sialytransferase 1 (ST8Sia1), and the consequently promoted inflammatory processes. Additionally, the expression of N-cadherin, Zinc finger protein SNAI2 (SLUG), and matrix metallopeptidase 9 (MMP-9) were elevated, but that of E-cadherin, which is known to be epithelial, was reduced. Moreover, the inhibition of ganglioside GD3 expression by the siRNA or exogenous treatment of neuraminidase 3 (NEU 3) led to the suppression of inflammation and EMT. These results suggest that gangliosides may play an important role in prevention and therapy for inflammation and EMT.


Engineering homoeologs provide a fine scale for quantitative traits in polyploid

August 2023

·

89 Reads

·

3 Citations

Plant Biotechnology Journal

Plant Biotechnology Journal

Numerous staple crops exhibit polyploidy and are difficult to genetically modify. However, recent advances in genome sequencing and editing have enabled polyploid genome engineering. The hexaploid black nightshade species Solanum nigrum has immense potential as a beneficial food supplement. We assembled its genome at the scaffold level. After functional annotations, we identified homoeologous gene sets, with similar sequence and expression profiles, based on comparative analyses of orthologous genes with close diploid relatives Solanum americanum and S. lycopersicum. Using CRISPR-Cas9-mediated mutagenesis, we generated various mutation combinations in homoeologous genes. Multiple mutants showed quantitative phenotypic changes based on the genotype, resulting in a broad-spectrum effect on the quantitative traits of hexaploid S. nigrum. Furthermore, we successfully improved the fruit productivity of Boranong, an orphan cultivar of S. nigrum suggesting that engineering homoeologous genes could be useful for agricultural improvement of polyploid crops.


Antifungal activity of CP4-EPSPS protein against fungal strains.
Potent Antifungal Functions of a Living Modified Organism Protein, CP4-EPSPS, against Pathogenic Fungal Cells

May 2023

·

30 Reads

·

1 Citation

Molecules

Various proteins introduced into living modified organism (LMO) crops function in plant defense mechanisms against target insect pests or herbicides. This study analyzed the antifungal effects of an introduced LMO protein, 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS) from Agrobacterium sp. strain CP4 (CP4-EPSPS). Pure recombinant CP4-EPSPS protein, expressed in Escherichia coli, inhibited the growth of human and plant fungal pathogens (Candida albicans, C. tropicalis, C. krusei, Colletotrichum gloeosporioides, Fusarium solani, F. graminearum, and Trichoderma virens), at minimum inhibitory concentrations (MICs) that ranged from 62.5 to 250 µg/mL. It inhibited fungal spore germination as well as cell proliferation on C. gloeosporioides. Rhodamine-labeled CP4-EPSPS accumulated on the fungal cell wall and within intracellular cytosol. In addition, the protein induced uptake of SYTOX Green into cells, but not into intracellular mitochondrial reactive oxygen species (ROS), indicating that its antifungal action was due to inducing the permeability of the fungal cell wall. Its antifungal action showed cell surface damage, as observed from fungal cell morphology. This study provided information on the effects of the LMO protein, EPSPS, on fungal growth.



Fig. 1 The effects of 7,8-dihydroxyflavone (7,8-DHF) on viability in malignant melanoma cells and HaCaT cells. A Chemical structure of 7,8-DHF. B The inhibitory effect of 7,8-DHF on the proliferation of melanoma cells. SK-MEL-2 (1.0 × 10 3 cells/well), G-361 (1.0 × 10 3 cells/well), and non-tumoral HaCaT cells (1.0 × 10 3 cells/well) were treated with different concentrations of 7,8-DHF for 24 and 48 h (hr). The viability was measured using an MTS assay kit. C Morphological changes in 7,8-DHF-treated (24 h) SK-MEL-2, G-361, and HaCaT cells and the corresponding untreated controls. Data represented the mean ± standard deviation (SD) of three independent experiments. ( * p < 0.05)
Fig. 2 Apoptotic effects induced by 7,8-DHF in malignant melanoma cells. SK-MEL-2, G-361, and HaCaT cells were maintained without 7,8-DHF (control group) or with different concentrations of 7,8-DHF (24 h). A Fluorescence microscopy images (original magnification: × 200) of nuclear staining with DAPI. White arrows represent DNA fragmentation and nuclear condensation. B Quantification of DNA fragmentation and condensation in melanoma cells. C Quantitative detection of Annexin-V/7-ADD positive cells was performed using a Muse ™ Cell analyzer. SK-MEL-2 (3.1 × 10 5 cells/well) and G-361 (4.3 × 10 5 cells/well) cells were treated with 7,8-DHF (100, 150, 200, 250, and 300 µM, 24 h). Cells stained with Annexin-V only were defined as early apoptotic, whereas Annexin-V and 7-ADD double-stained cells were defined as late apoptotic. D The percentage of the total apoptotic cell population was quantified. Data were presented as mean percentage levels ± SD (n = 3, * p < 0.05)
Fig. 3 Anti-proliferative and apoptotic effects induced by 7,8-DHF in malignant melanoma cells. A SK-MEL-2 (1.0 × 10 5 cells/well) and G-361 cells (1.0 × 10 5 cells/well) were incubated without 7,8-DHF (control group) or with 7,8-DHF (100, 200, and 300 µM for 24 h). Cells were then washed with DPBS, fixed in 70% ethanol overnight, stained with Muse ™ Cell-Cycle reagent, and finally analyzed for DNA content using a Muse ™ Cell analyzer. B The results of cell cycle population in SK-MEL-2 and G-361 cells were quantified. Data represented the mean ± SD (n = 3, * p < 0.05). C SK-MEL-2 and G-361 cells were treated with different concentrations of 7,8-DHF (100, 200, and 300 µM for 24 h). Whole cells extracts were prepared, separated on SDS-PAGE, and transferred onto blotting membranes. Western blot analysis was performed using p53, p21, Cyclin B, CDK1, MCL-1, and Survivin antibodies. D SK-MEL-2 and G-361 cells were treated with 7,8-DHF (100, 200, and 300 µM for 24 h). The lysate samples were subjected to western blot analysis with BID, BAX, BCL-X L, Caspase 3 (CASP3), cleaved-CASP3, poly (ADPribose) polymerase (PARP), and cleaved-PARP antibodies. β-actin was used as a loading control. The results shown in C, D were representative of at least three independent experiments
Fig. 4 Changes in the mitochondrial membrane potential of malignant SK-MEL-2 and G-361 melanoma cells. A Mitochondrial membrane depolarization by using JC-1 staining was determined through the immunofluorescence microscopy. Melanoma SK-MEL-2 (1.0 × 10 5 cells/ well) and G-361 cells (1.0 × 10 5 cells/well) were treated with 7,8-DHF (100 and 200 µM) for 24 h and mitochondrial membrane depolarization was found to increase in a dose-dependent manner. B The mitochondrial apoptotic process in SK-MEL-2 and G-361 cells was determined by western blot analysis of mitochondrial and cytosolic protein levels. The mitochondrial fraction was confirmed using the mitochondrial marker MTCO1, and the cytosolic fraction was confirmed using β-tubulin. 7,8-DHF induced Cytochrome c release and Bax expression. 7,8-DHF decreased BID, BCL-X L, Caspase 9, CASP3, and PARP expression time-dependently. Cyto, cytosolic fraction; Mito, mitochondrial fraction
Fig. 5 7,8-DHF significantly inhibited the metastatic ability of SK-MEL-2 and G-361 cells. A Effects of 7,8-DHF on the migration of melanoma cell lines. SK-MEL-2 (5.0 × 10 5 cells/well) and G-371 cells (5.0 × 10 5 cells/well) were treated with 7,8-DHF (200 µM). Melanoma cells were scratched using sterile pipette tips and then incubated with or without (control group) 7,8-DHF. Samples were observed under a light microscope at 24, 48, and 72 h. B The area covered with cells was quantified. Red square and blue square represent SK-MEL-2 and G-361 cells, respectively. All data were presented as the mean ± SD (n = 3, * p < 0.05)
7,8-Dihydroxyflavone induces mitochondrial apoptosis and down-regulates the expression of ganglioside GD3 in malignant melanoma cells

March 2023

·

43 Reads

·

4 Citations

Discover Oncology

Malignant melanoma is a skin cancer with poor prognosis and high resistance to conventional treatment. 7,8-dihydroxyflavone (7,8-DHF) has shown anti-carcinogenic, anti-inflammatory, anti-oxidant, and pharmacological effects in several types of cancer. However, the relationship between ganglioside expression and the anti-cancer effects of 7,8-DHF in melanoma is not fully understood. In the present study, 7,8-DHF exhibits specific anti-proliferation, anti-migration, and G2/M phase cell-cycle arrest effects on both melanoma cancer cell lines, and induces mitochondrial dysfunction and apoptosis, making it a potent candidate for anti-melanoma treatment. Furthermore, we confirmed that 7,8-DHF significantly reduces the expression levels of ganglioside GD3 and its synthase, which are known to be closely involved in carcinogenesis. Taken together, our findings suggest that 7,8-DHF may be a potent anti-cancer drug candidate for the treatment of malignant melanoma.


7,8-Dihydroxyflavone induces mitochondrial apoptosis and down-regulates the expression of ganglioside GD3 in malignant melanoma cells

June 2022

·

5 Reads

Malignant melanoma is one of the most progressive skin cancers, with a poor prognosis, various side effects, and high resistance to conventional treatment. Recently, 7,8-dihydroxyflavone (7,8-DHF), a polyphenolic flavonoid derived from certain plants, has been studied for its anti-carcinogenic, anti-inflammatory, antioxidant, and pharmacological effects in several types of cancer. Ganglioside, a modulator of diverse cell signals on the microdomain of the surface of the cell membrane, is closely involved in various cancers such as neuroblastoma. However, the correlation between ganglioside expression and the anti-cancer effects of 7,8-DHF in malignant melanoma remains unclear. In this study, 7,8-DHF showed potential as an anti-cancer agent through specific anti-proliferation, anti-oxidant, anti-migration, pro-apoptotic, and G2/M phase cell cycle arrest effects on SK-MEL-2 and G-361 melanoma cells. In contrast, 7,8-DHF did not induce cytotoxicity in non-tumoral epidermal HaCaT cells. Additionally, we confirmed for the first time that 7,8-DHF significantly reduces the expression levels of ganglioside GD3, which is closely involved in carcinogenesis, in both melanoma cancer cell lines using high-performance thin-layer chromatography analysis. Taken together, our findings suggest that 7,8-DHF might be a potent anticancer drug candidate for the treatment of malignant melanoma.


7,8-Dihydroxyflavone induces mitochondrial apoptosis and down-regulates the expression of ganglioside GD3 in malignant melanoma cells

June 2022

·

8 Reads

Malignant melanoma is one of the most progressive skin cancers, with a poor prognosis, various side effects, and high resistance to conventional treatment. Recently, 7,8-dihydroxyflavone (7,8-DHF), a polyphenolic flavonoid derived from certain plants, has been studied for its anti-carcinogenic, anti-inflammatory, antioxidant, and pharmacological effects in several types of cancer. Ganglioside, a modulator of diverse cell signals on the microdomain of the surface of the cell membrane, is closely involved in various cancers. However, the correlation between ganglioside expression and the anti-cancer effects of 7,8-DHF in malignant melanoma remains unclear. In this study, 7,8-DHF showed potential as an anti-cancer agent through specific anti-proliferation, anti-oxidant, anti-migration, pro-apoptotic, and G2/M phase cell cycle arrest effects on SK-MEL-2 and G-361 melanoma cells. In contrast, 7,8-DHF did not induce cytotoxicity in non-tumoral epidermal HaCaT cells. Additionally, we confirmed for the first time that 7,8-DHF significantly reduces the expression levels of ganglioside GD3, which is closely involved in carcinogenesis, in both melanoma cancer cell lines using high-performance thin-layer chromatography analysis. Taken together, our findings suggest that 7,8-DHF might be a potent anticancer drug candidate for the treatment of malignant melanoma.


Narrow lpa1 Metaxylems Enhance Drought Tolerance and Optimize Water Use for Grain Filling in Dwarf Rice

May 2022

·

209 Reads

·

6 Citations

Rice cultivation needs extensive amounts of water. Moreover, increased frequency of droughts and water scarcity has become a global concern for rice cultivation. Hence, optimization of water use is crucial for sustainable agriculture. Here, we characterized Loose Plant Architecture 1 (LPA1) in vasculature development, water transport, drought resistance, and grain yield. We performed genetic combination of lpa1 with semi-dwarf mutant to offer the optimum rice architecture for more efficient water use. LPA1 expressed in pre-vascular cells of leaf primordia regulates genes associated with carbohydrate metabolism and cell enlargement. Thus, it plays a role in metaxylem enlargement of the aerial organs. Narrow metaxylem of lpa1 exhibit leaves curling on sunny day and convey drought tolerance but reduce grain yield in mature plants. However, the genetic combination of lpa1 with semi-dwarf mutant (dep1-ko or d2) offer optimal water supply and drought resistance without impacting grain-filling rates. Our results show that water use, and transports can be genetically controlled by optimizing metaxylem vessel size and plant height, which may be utilized for enhancing drought tolerance and offers the potential solution to face the more frequent harsh climate condition in the future.


Figure 2. SDS-PAGE analysis and verification of high-abundance proteins in MSM. (a) SDS-PAGE and silver staining: prestained protein size marker (lane M), secretomes from cultured medium of active MPs (lane MSM), and sample picking from SDS-PAGE gel spots for peptide mass fingerprinting (PMF) analysis (lane Picks). N.D., not detected. (b) Quantification of protein bands from SDS-PAGE gels using Image J software. Data are presented as mean fold changes ± SD (n = 3; #, more than 10-fold increase versus the control, p < 0.05; ##, more than 20-fold increase versus the control, p < 0.05). (c) Two clusters were identified using the DAVID functional annotation clustering tool. Annotated cluster represents a kappa value > 0.35 and overlap = 3. Similarity scores ranged from high (>1) to low enrichment (<0.25). (d) Illustration of identified proteins analyzed by gene ontology tool. (e) PMF spectrum of NME1. Spectral masses (in mass per charge unit, m/z) obtained by MALDI-TOF MS were analyzed by bioinformatics using the Mascot and Profound search engines. The x-axis represents the mass-to-charge ratio (m/z), and the y-axis represents the relative abundance. The probability-based Mowse scores were obtained using the Mascot search engine. Among predicted proteins with differential Mowse scores shown as multiple bars on the x-axis, only proteins with Mowse scores > 68 were considered significant, which was 142 (p < 0.05) for NME1 protein. Red marks indicate peptides identified by mass fingerprinting that match calculated molecular weights. All protein identifications are provided in Supplementary Figure S2 and Table S2.
Figure 4. Identification of hNME1 as a novel pST8SIA1-binding protein using IP and pull-down assay. (a) NME1 interacts with ST8SIA1. U937 cells were lysed under non-denaturing conditions, and 500 μg of total lysate protein was subjected to IP with α-NME1. Precipitated proteins were resolved by SDS-PAGE, and IB analysis probed for ST8SIA1 or NME1 as indicated. Arrows indicate immunoreactive bands; the precipitated ST8SIA1 band is denoted with an asterisk. Beads, beads only (IgG); L/C, light chain. (b) The rpST8SIA1 constructs. GST fusion proteins were generated by cloning PCRamplified ST8SIA1 sequences into the pEX-N-GST expression vector. Domain structure of the full-length GST-ST8SIA1 protein and the various deletion mutants used. X1-3, transcript variants 1-3; P1-3, partial transcripts 1-3; aa, amino acids. (c) Equivalent amounts of purified recombinant GST, GST-ST8SIA1, or GST-ST8SIA1 deletion mutants were analyzed by SDS-PAGE followed by IB analysis with the specific α-GST antibody. (d) GST pull-down assay on purified-His-rhNME1 incubated with equivalent amounts of GST, GST-X1, GST-X2, GST-X3, GST-P1, GST-P2, and GST-P3. Results were analyzed by SDS-PAGE followed by IB analysis with the specific α-GST and α-His antibody. The ST8SIA1 domains bound by NME1 are denoted with an asterisk. (e) GST pull-down assay on His-rhNME1 or purified-His-tagged recombinant porcine NME1 incubated with equivalent amounts of GST and GST-X1 beads. Results were analyzed by SDS-PAGE followed by
Figure 5. Immunocytochemistry of mp AD-MSCs with and without rhNME1 treatment. (a) Co-localization of endogenous ST8SIA1 in the Golgi apparatus in mp AD-MSCs or endogenous ST8SIA1 with exogenous rhNME1. Immunofluorescence microscopy results show mp AD-MSCs fixed and stained with antibodies against ST8SIA1 (green), cis-Golgi marker GM130 (red), and ectopic His-tagged rhNME1 (red). DAPI was used for nuclear counter staining. Overlapping staining (yellow) demonstrates the co-localization of the two proteins, and white squares indicate enlarged merged images. (b) Reduction in ST8SIA1 expression by rhNME1, rsNME1, and MSM treatment in mp AD-MSCs or NI-mp AD-MSCs. RT-PCR (left panel) and Western blots (right panel) for ST8SIA1 protein in rhNME1 or rsNME1-treated NI-mp AD-MSCs. Only rhNME1 (1, 2, and 4 ng/mL), not rsNME1, dose-dependently significantly decreased ST8SIA1 protein levels in NImp AD-MSCs; neither rhNME1 nor rsNME1 suppressed ST8SIA1 mRNA in NI-mp AD-MSCs. ACTB was used as a loading control. (c) ST8SIA1 is regulated by the proteasome and rhNME1. Western blot analysis determined the effects of rhNME1, MG132, and cycloheximide on ST8SIA1 protein accumulation. Protein extracts were prepared from NI-mp ADMSCs treated with rhNME1 (4 ng/mL) or MG132 (10 uM), cycloheximide (150 nM), or a combination of the two. Equal protein loading was confirmed with ACTB antibody. (d) The knockdown of ST8SIA1 siRNA. ST8SIA1-specific siST8SIA1 or siCon was transfected into mp AD-MSCs. After transfection with siST8SIA1 (50 nM) for 48 h, ST8SIA1 expression was evaluated using HPTLC, RT-PCR, and Western blotting. ACTB was used as a loading control. All siRNA sequence information is provided in Supplementary Figure S4 and Table S3. #, siRNA number. (e) Phase-contrast images of transfected NI-mp AD-MSCs with siCon or siST8SIA1 (#4). Arrowheads indicate enlarged images. (f) Variations in the cell count of NI-mp AD-MSCs were quantified for NI-mp AD-MSCs transfected with siCon or siST8SIA1#4 (si#4). Data are presented as mean percentage levels ± SD (n = 3; * p < 0.05).
The Potential Role of Human NME1 in Neuronal Differentiation of Porcine Mesenchymal Stem Cells: Application of NB-hNME1 as a Human NME1 Suppressor

November 2021

·

50 Reads

·

3 Citations

International Journal of Molecular Sciences

This study aimed to investigate the effects of the human macrophage (MP) secretome in cellular xenograft rejection. The role of human nucleoside diphosphate kinase A (hNME1), from the secretome of MPs involved in the neuronal differentiation of miniature pig adipose tissue-derived mesenchymal stem cells (mp AD-MSCs), was evaluated by proteomic analysis. Herein, we first demonstrate that hNME1 strongly binds to porcine ST8 alpha-N-acetyl-neuraminide alpha-2,8-sialyltransferase 1 (pST8SIA1), which is a ganglioside GD3 synthase. When hNME1 binds with pST8SIA1, it induces degradation of pST8SIA1 in mp AD-MSCs, thereby inhibiting the expression of ganglioside GD3 followed by decreased neuronal differentiation of mp AD-MSCs. Therefore, we produced nanobodies (NBs) named NB-hNME1 that bind to hNME1 specifically, and the inhibitory effect of NB-hNME1 was evaluated for blocking the binding between hNME1 and pST8SIA1. Consequently, NB-hNME1 effectively blocked the binding of hNME1 to pST8SIA1, thereby recovering the expression of ganglioside GD3 and neuronal differentiation of mp AD-MSCs. Our findings suggest that mp AD-MSCs could be a potential candidate for use as an additive, such as an immunosuppressant, in stem cell transplantation.


Citations (73)


... [5][6][7] CRISPR-Cas9 technology has rapidly become a very effective method for genome editing, and is thus widely used in animals and plants. [8][9][10] The system consists of Cas9, Cas1, Cas2, and Csn1 proteins as well as CRISPR RNA (crRNA) and trans-activating CRISPR RNA (tracrRNA). 11,12 Cas9 can form a complex with synthetic single guide RNA (sgRNA), the sgRNA is formed by fusion of crRNA and trans-activated tracrRNA, the sgRNA guides Cas9 to recognize and cleave the target DNA. ...

Reference:

Optimization of CRISPR/Cas9 system in Eustoma grandiflorum
Engineering homoeologs provide a fine scale for quantitative traits in polyploid
  • Citing Article
  • August 2023

Plant Biotechnology Journal

Plant Biotechnology Journal

... Yeast fungi were cultured overnight in YPD broth, and mold spores were collected from 4-day-old cultures grown on PD agar plates using 0.08% Triton X-100. Fungal suspensions in Buffer A, or 25 mM MES buffer with 150 mM NaCl (pH 5.5) containing 20% of the appropriate media (2 × 10 4 cells or spores/mL), were added to serially diluted AtTCP21 proteins and melittin peptides in 96-well plates [19]. After 24 h incubation at 28 • C, 10 µL of cells was dropped on PDA agar, and cell/mycelial growth was measured at 595 nm using an ELISA reader (Molecular Devices M5, Sunnyvale, CA, USA). ...

Potent Antifungal Functions of a Living Modified Organism Protein, CP4-EPSPS, against Pathogenic Fungal Cells

Molecules

... The examination of ganglioside expression in melanoma cells revealed elevated levels of both ganglioside GD3 and GM3, consistent with prior reports [47,48]. Quercetin treatment effectively decreased the expression levels of these gangliosides, aligning with findings using other flavonoid drugs [49]. The data suggest ganglioside GM3 and GD3 as potential specific markers for melanoma cells, potentially serving as therapeutic targets in these cancers. ...

7,8-Dihydroxyflavone induces mitochondrial apoptosis and down-regulates the expression of ganglioside GD3 in malignant melanoma cells

Discover Oncology

... From this perspective, the search for advances in breeding programs aimed at resistance to drought stress in agricultural crops is a path that should be pursued with greater emphasis. Breeding programs have been working with different crops and mechanisms to investigate resistance to drought stress, including gene expression, genetically modified organisms, plant hormones, biostimulants, and the identification of genes that confer resistance in crops such as corn, soybean, cotton, rice, wheat, common bean, sugarcane, and tomato, among others [1,4,[7][8][9][10][11][12][13][14][15][16][17]. ...

Narrow lpa1 Metaxylems Enhance Drought Tolerance and Optimize Water Use for Grain Filling in Dwarf Rice
Frontiers in Plant Science

Frontiers in Plant Science

... Cho et al. demonstrated the critical role of ganglioside GD3 in the neuronal differentiation of pig adipose stem cells. Specifically, enzyme ganglioside synthase inhibition causes the blocking of ganglioside GD3 expression followed by reduced neuronal differentiation of ADSCs [90]. Regarding the adipogenic differentiation of MSCs, GM3 is the highest in adipocytes. ...

The Potential Role of Human NME1 in Neuronal Differentiation of Porcine Mesenchymal Stem Cells: Application of NB-hNME1 as a Human NME1 Suppressor

International Journal of Molecular Sciences

... By further investigating the molecular mechanisms of MID-269aa-induced MARCKS translocation, we found that calmodulin was another downstream target that directly interacted with TRIM1-269aa. CaM can be encoded by a member of the CALM gene family (CALM1, CALM2, and CALM3), which is a Ca 2+ -binding protein with high affinity for the MARCKS protein [63]. Previous studies have reported that calmodulin shares the same MARCKS PSD that serves as the site for PKC phosphorylation [64], and the combination of MARCKS PSD with calmodulin leads to its dissociation from the plasma membrane and subsequent modulation of cellular processes such as cytoskeletal reorganization, membrane trafficking and activation of the PI3K/ AKT/mTOR pathway [65]. ...

Targeting CALM2 Inhibits Hepatocellular Carcinoma Growth and Metastasis by Suppressing E2F5-mediated Cell Cycle Progression
  • Citing Article
  • March 2021

Anticancer Research

... KRAS is one of the main driving genes in pancreatic cancer. More than 90% of pancreatic cancer patients have KRAS carcinogenic point mutations, which constitutively activate the RAS signaling pathway [12][13][14]. The RAS signal transduction regulates multiple cellular biological processes in the pancreatic ductal adenocarcinoma, including the proliferation, migration, metabolism, and autophagy of the cells [15]. ...

Establishment and Characterization of Immortalized Miniature Pig Pancreatic Cell Lines Expressing Oncogenic K-RasG12D

International Journal of Molecular Sciences

... However, their poor engraftment and low survival upon transplantation led to the re-evaluation of the mechanisms underlying beneficial effects of MSC in a context-specific manner 29 . Of the various pleiotropic functions, immunomodulatory properties make MSC a promising strategy for treating immune-mediated disorders 30 . On the other hand, since immunosuppressive potential of MSC can be enhanced by culture or environment engineering, great efforts have been made to improve the therapeutic efficacy of MSC 31 . ...

Application of Mesenchymal Stem Cells in Inflammatory and Fibrotic Diseases

International Journal of Molecular Sciences

... The spleen is one of the biggest organs in human/animal body. If the ratio of spleen to body wt% augmented, there are more immune cells in the spleen, that is to say, the immune response is more serious [66]. After the IMQ treatment for 6 days, the percentage of spleen/body weight (1.05 ± 0.37) in the IMQ only group was more than 2 times higher than that in the normal mice group (0.5 ± 0.19) after 6 days of administration, indicating that the number of spleen cells increased ( Fig. 12A and B). ...

Therapeutic anti-psoriatic effects of myeloid-derived suppressor cells in combination with systemic tacrolimus (FK-506) in an imiquimod-induced mouse model of psoriasis
  • Citing Article
  • September 2020

International Immunopharmacology

... GSLs play a crucial role in the repair and differentiation processes of articular cartilage [50,59]. These processes are essential for addressing cartilage defects, commonly observed in osteoarthritis. ...

Ganglioside GM3 Up-Regulate Chondrogenic Differentiation by Transform Growth Factor Receptors

International Journal of Molecular Sciences