Objective: To establish a C26 cell strain stably expressing folate receptor α (FRα) for the subsequent study of FRα DNA vaccine.
Methods: C26 cells were transfected with previously constructed recombinant eukaryotic expressing vector pcDNA3.1-FRα by LipofectamineTM2000. Afterwards the cells were subjected to G418 (500 mg/mL) selection to get G418 resistant cells. And then single cell cloning was performed to generate monoclonal cell strain. The gene and protein expression levels of FRα of the monoclonal cell strain were further analyzed by reverse transcription PCR (RT-PCR) and fluorescence microscopy. The cells were cultured for several generations, and the mRNA expression of FRα was analyzed by RT-PCR at different generations to determine whether the transfected gene was stable or not during cell passaging.
Results: After transfection, G418 selection and single cell cloning, the monoclonal cell strain were established and proved to be able to express FRα mRNA and protein and keep the stability of FRα expression after several generations.
Conclusion: We have established the cell strain stably expressing FRα, which offer a tool to evaluate the effect of DNA vaccine based on FRα.
Download full-text PDF |
Source |
|---|
Publication Analysis
Top Keywords
Similar Publications
Background: Small, soluble oligomers, rather than mature fibrils, are the major neurotoxic agents in Alzheimer's disease (AD). In the last few years, Aprile and co-workers designed and purified a single-domain antibody (sdAb), called DesAb-O, with high specificity for Aβ oligomeric conformers. Recently, Cascella and co-workers showed that DesAb-O can selectively detect synthetic Aβ oligomers both in vitro and in cultured cells, neutralizing their associated neuronal dysfunction.
View Article and Find Full Text PDFBackground: Human pluripotent stem cell (hPSC)-derived brain organoids patterned towards the cerebral cortex are valuable models of interactions occurring in vivo in cortical tissue. We and others have used these cortical organoids to model dominantly inherited FTD-tau. While these studies have provided essential insights, cortical organoid models have yet to reach their full potential.
View Article and Find Full Text PDFDrug Development.
Alzheimers Dement
December 2024
Division of Neurodegenerative Disorders, St. Boniface Hospital Albrechtsen Research Centre, Winnipeg, MB, Canada.
Background: Alzheimer's disease (AD) is a neurodegenerative disorder primarily associated with aging, but manifests as a complex interplay of multiple factors. Decline in sex-hormones, particularly 17-beta estradiol, is linked to the aging process. The risk for onset of AD significantly increases with aging and loss of estradiol.
View Article and Find Full Text PDFGeneration of Retinal Ganglion Cells from Reprogrammed Keratocytes of Non-Glaucoma and Glaucoma Donors.
Curr Protoc
January 2025
Department of Ophthalmology, Indiana University School of Medicine, Indianapolis, Indiana.
Human induced pluripotent stem cell (hiPSC)-based disease modeling can be successfully recapitulated to mimic disease characteristics across various human pathologies. Glaucoma, a progressive optic neuropathy, primarily affects the retinal ganglion cells (RGCs). While multiple groups have successfully generated RGCs from non-diseased hiPSCs, producing RGCs from glaucomatous human samples holds significant promise for understanding disease pathology by revealing patient-specific disease signatures.
View Article and Find Full Text PDFExploring vimentin's role in breast cancer via PICK1 alternative polyadenylation and the miR-615-3p-PICK1 interaction.
Biofactors
January 2025
Department of Oncology, Shanghai East Hospital, Tongji University School of Medicine, Tongji University, Shanghai, People's Republic of China.
Breast cancer continues to be a major health issue for women worldwide, with vimentin (VIM) identified as a crucial factor in its progression due to its role in cell migration and the epithelial-to-mesenchymal transition (EMT). This study focuses on elucidating VIM's regulatory mechanisms on the miR-615-3p/PICK1 axis. Utilizing the 4T1 breast cancer cell model, we first used RNA-seq and proteomics to investigate the changes in the APA of PICK1 following VIM knockout (KO).
View Article and Find Full Text PDFWant AI Summaries of new PubMed Abstracts delivered to your In-box?
Enter search terms and have AI summaries delivered each week - change queries or unsubscribe any time!