scholarly journals Expression of Recombinant Human Octamer-Binding Transcription Factor 4 in Rice Suspension Cells

2021 ◽  
Vol 22 (3) ◽  
pp. 1409
Author(s):  
Li-Fen Huang ◽  
Desyanti Saulina Sinaga ◽  
Chia-Chun Tan ◽  
Shu-Ju Micky Hsieh ◽  
Chi-Hung Huang
Keyword(s):  
Stem Cells ◽  
Transcription Factor ◽  
Recombinant Protein ◽  
Transgenic Rice ◽  
Expression System ◽  
Suspension Cells ◽  
Sugar Starvation ◽  
Oct4 Protein ◽  
Octamer Binding Transcription Factor ◽  
Transcription Factor 4

The rice cell suspension culture system is a good way to produce recombinant human proteins, owing to its high biosafety and low production cost. Human Octamer-binding Transcription Factor 4 (Oct4) is a fundamental transcription factor responsible for maintaining human pluripotent embryonic stem cells. Recombinant Oct4 protein has been used to induce pluripotent stem cells. In this study, recombinant Oct4 proteins are produced via a sugar starvation-inducible αAmy3/RAmy3D promoter–signal peptide-based rice recombinant protein expression system. Oct4 mRNAs accumulate in the transgenic rice suspension cells under sugar starvation. The Oct4 recombinant protein is detected in the transgenic rice suspension cells, and its highest yield is approximately 0.41% of total cellular soluble proteins after one day of sugar starvation. The rice cell-synthesized recombinant human Oct4 protein show DNA-binding activity in vitro, which implies that the protein structure is correct for enabling specific binding to the target DNA motif.

scholarly journals In silico identification of natural product inhibitors against Octamer-binding transcription factor 4 (Oct4) to impede the mechanism of glioma stem cells

PLoS ONE ◽  
2021 ◽  
Vol 16 (10) ◽  
pp. e0255803
Author(s):  
Chirasmita Nayak ◽  
Sanjeev Kumar Singh
Keyword(s):  
Stem Cells ◽  
Transcription Factor ◽  
Natural Product ◽  
Binding Free Energy ◽  
Dynamics Simulation ◽  
Strong Interactions ◽  
Energy Calculation ◽  
Specific Domain ◽  
Octamer Binding Transcription Factor ◽  
Transcription Factor 4

Octamer-binding transcription factor 4 (Oct4) is a core regulator in the retention of stemness, invasive, and self-renewal properties in glioma initiating cells (GSCs) and its overexpression inhibits the differentiation of glioma cells promoting tumor cell proliferation. The Pit-Oct-Unc (POU) domain comprising POU-specific domain (POUS) and POU-type homeodomain (POUHD) subdomains is the most critical part of the Oct4 for the generation of induced pluripotent stem cells from somatic cells that lead to tumor initiation, invasion, posttreatment relapse, and therapeutic resistance. Therefore, the present investigation hunts for natural product inhibitors (NPIs) against the POUHD domain of Oct4 by employing receptor-based virtual screening (RBVS) followed by binding free energy calculation and molecular dynamics simulation (MDS). RBVS provided 13 compounds with acceptable ranges of pharmacokinetic properties and good docking scores having key interactions with the POUHD domain. More Specifically, conformational and interaction stability analysis of 13 compounds through MDS unveiled two compounds ZINC02145000 and ZINC32124203 which stabilized the backbone of protein even in the presence of linker and POUS domain. Additionally, ZINC02145000 and ZINC32124203 exhibited stable and strong interactions with key residues W277, R242, and R234 of the POUHD domain even in dynamic conditions. Interestingly, ZINC02145000 and ZINC32124203 established communication not only with the POUHD domain but also with the POUS domain indicating their incredible potency toward thwarting the function of Oct4. ZINC02145000 and ZINC32124203 also reduced the flexibility and escalated the correlations between the amino acid residues of Oct4 evidenced by PCA and DCCM analysis. Finally, our examination proposed two NPIs that can impede the Oct4 function and may help to improve overall survival, diminish tumor relapse, and achieve a cure not only in deadly disease GBM but also in other cancers with minimal side effects.

scholarly journals Subpopulations of miniature pig mesenchymal stromal cells with different differentiation potentials differ in the expression of octamer-binding transcription factor 4 and sex determining region Y-box 2

2020 ◽  
Vol 33 (3) ◽  
pp. 515-524
Author(s):  
Ryounghoon Jeon ◽  
Sungjo Park ◽  
Sung-Lim Lee ◽  
Gyu-Jin Rho
Keyword(s):  
Transcription Factor ◽  
Stromal Cells ◽  
Antigen Expression ◽  
Miniature Pig ◽  
Proliferative Capacity ◽  
Differentiation Potential ◽  
Sox2 Expression ◽  
Surface Antigen Expression ◽  
Octamer Binding Transcription Factor ◽  
Transcription Factor 4

Objective: Human mesenchymal stromal cells (MSCs) exhibit variable differentiation potential and can be divided accordingly into distinct subpopulations whose ratios vary with donor age. However, it is unknown whether the same is true in pigs. This study investigated MSC subpopulations in miniature pig and compared their characteristics in young (2 to 3 months) and adult (27 to 35 months) pigs.Methods: Osteogenic, chondrogenic, and adipogenic capacity of isolated MSCs was evaluated by von Kossa, Alcian blue, and oil red O staining, respectively. Cell surface antigen expression was determined by flow cytometry. Proliferative capacity was assessed with the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. Expression of marker genes was detected by quantitative real-time polymerase chain reaction.Results: Porcine MSCs comprised cells with trilineage and bilineage differentiation potential (tMSCs and bMSCs, respectively) and non-differentiating stromal cells (NDSCs). The tMSC and bMSC fractions were smaller in adult than in young pigs (63.0% vs 71.2% and 11.6% vs 24.0%, respectively, p<0.05); NDSCs showed the opposite trend (25.4% vs 4.8%; p<0.05). Subpopulations showed no differences in morphology, cell surface antigen expression, or proliferative capacity, but octamer-binding transcription factor 4 (OCT4) expression was higher in tMSCs than in bMSCs and NDSCs (p<0.05), whereas sex determining region Y-box 2 (SOX2) expression was higher in tMSCs and bMSCs than in NDSCs (p<0.05). Aging had no effect on these trends.Conclusion: Porcine MSCs comprise distinct subpopulations that differ in their differentiation potential and OCT4 and SOX2 expression. Aging does not affect the characteristics of each subpopulation but alters their ratios.

Distribution pattern of cytoplasmic organelles, spindle integrity, oxidative stress, octamer-binding transcription factor 4 (Oct4) expression and developmental potential of oocytes following multiple superovulation

2016 ◽  
Vol 28 (12) ◽  
pp. 2027 ◽  
Author(s):  
Guruprasad Kalthur ◽  
Sujith Raj Salian ◽  
Ramya Nair ◽  
Jemey Mathew ◽  
Satish Kumar Adiga ◽  
...  
Keyword(s):  
Oxidative Stress ◽  
Transcription Factor ◽  
Oocyte Quality ◽  
Hatching Rate ◽  
Developmental Potential ◽  
Oct4 Expression ◽  
Cytoplasmic Organelles ◽  
Serum Gonadotropin ◽  
Octamer Binding Transcription Factor ◽  
Transcription Factor 4

The aim of the present study was to determine the effects of repeated superovulation on oocyte quality and embryo developmental potential. Female Swiss albino mice were injected with 5 IU pregnant mare’s serum gonadotropin followed 48 h by 10 IU human chorionic gonadotropin. Mice were superovulated up to four times with a gap of 7 days between each superovulation cycle. Ovarian weight increased significantly with an increasing number of superovulation cycles. Although the first stimulation cycle resulted in a threefold increase in the number of oocytes, the number of oocytes decreased gradually after subsequent stimulations. Increased cytoplasmic fragmentation, abnormal mitochondrial distribution, aggregation of Golgi apparatus, spindle damage, increased intracellular oxidative stress and a decrease in expression of octamer-binding transcription factor 4 (Oct4) expression were observed in these oocytes. Further, embryos derived from mice subjected to multiple stimulation cycles exhibited a low blastocyst rate, decreased hatching rate and increased apoptosis in blastocysts. In conclusion, the present study demonstrates that repeated superovulation adversely affects mouse oocyte quality by altering the distribution of cytoplasmic organelles, increasing oxidative stress and decreasing Oct4 expression, resulting in poor developmental potential of the embryos.

scholarly journals Inhibition of C-terminal binding protein attenuates transcription factor 4 signaling to selectively target colon cancer stem cells

Cell Cycle ◽  
2014 ◽  
Vol 13 (22) ◽  
pp. 3506-3518 ◽  
Author(s):  
Jagrut Patel ◽  
Somesh Baranwal ◽  
Ian M Love ◽  
Nirmita J Patel ◽  
Steven R Grossman ◽  
...  
Keyword(s):  
Stem Cells ◽  
Colon Cancer ◽  
Transcription Factor ◽  
Cancer Stem Cells ◽  
Binding Protein ◽  
Colon Cancer Stem Cells ◽  
Transcription Factor 4
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