Creation of CD63-deficient HEK293 cell lines using a polycistronic CAS9/EGFP/HSVtk/PuroR expression vector

Matters ◽  
2016 ◽  
Author(s):  
Francis Fordjour ◽  
Shawn Owiredu ◽  
Hayley Muendlein ◽  
Jerry Plange ◽  
James Morrell ◽  
...  
Keyword(s):  
Cell Lines ◽  
Expression Vector ◽  
Hek293 Cell

Transient gene expression optimization and expression vector comparison to improve HIV-1 VLP production in HEK293 cell lines

2017 ◽  
Vol 102 (1) ◽  
pp. 165-174 ◽  
Author(s):  
Javier Fuenmayor ◽  
Laura Cervera ◽  
Sonia Gutiérrez-Granados ◽  
Francesc Gòdia
Keyword(s):  
Gene Expression ◽  
Cell Lines ◽  
Expression Vector ◽  
Hek293 Cell ◽  
Transient Gene Expression ◽  
Expression Optimization ◽  
Hiv 1

Lemon Juice Mediated Synthesis of 3-Substituted Quinazolin-4(3H)-Ones and their Pharmacological Evaluation

2020 ◽  
Vol 19 (16) ◽  
pp. 2001-2009 ◽  
Author(s):  
Malavattu G. Prasad ◽  
C. Vijaya Lakshmi ◽  
Naresh K. Katari ◽  
Sreekantha B. Jonnalagadda ◽  
Manojit Pal
Keyword(s):  
Cell Line ◽  
Cell Lines ◽  
Hek293 Cell ◽  
Glyoxylic Acid ◽  
Lemon Juice ◽  
Cytotoxic Agents ◽  
Diverse Range ◽  
Hek293 Cell Line ◽  
Three Component Reaction ◽  
Mcf 7

Background: Compounds containing the quinazoline-4(3H)-one framework constitute an important class of fused N-heterocycles that are found in more than 200 naturally occurring alkaloids. These compounds also show a diverse range of pharmacological activities including antitumor properties. This prompted us to explore a series of quinazolin-4-(3H)-one derivatives having no substituent at C-2 as potential cytotoxic agents. Objective: The objective of this study was to synthesize and evaluate 3-substituted quinazolin-4(3H)-one derivatives for their potential cytotoxic properties. Methods: A convenient method has been developed for the rapid synthesis of this class of compounds under a mild and non-hazardous reaction condition in good yields. The methodology involved a three-component reaction employing isatoic anhydride, amines and glyoxylic acid as reactants in the presence of lemon juice in PEG- 400 at room temperature (25-30ºC) under ultrasound irradiation. All the synthesized compounds were screened via an MTT assay for their potential cytotoxic properties in vitro using the cancerous cell lines e.g. A549, A2780, HepG2, K562, MCF-7 and HCT-116 and a non-cancerous HEK293 cell line. Results: Several compounds such as 3a, 3b, 3d, 3e and 3f showed promising growth inhibition against these cancer cell lines but no significant effects on HEK293 cell line. The IC50 values of these compounds were comparable to doxorubicin whereas 3f significantly induced apoptosis in MCF-7 cells that also was comparable to doxorubicin. Conclusion: An ultrasound-assisted MCR facilitated by lemon juice has been developed to synthesize 3- substituted quinazolin-4(3H)-one derivatives that could act as potential anticancer agents.

scholarly journals Affecting HEK293 Cell Growth and Production Performance by Modifying the Expression of Specific Genes

Cells ◽  
2021 ◽  
Vol 10 (7) ◽  
pp. 1667
Author(s):  
Laura Abaandou ◽  
David Quan ◽  
Joseph Shiloach
Keyword(s):  
Cell Line ◽  
Cell Lines ◽  
Hek293 Cell ◽  
Chinese Hamster ◽  
Production Performance ◽  
Cellular Processes ◽  
Hek293 Cell Line ◽  
Global Engineering ◽  
Genomic Tools ◽  
Serum Free Suspension

The HEK293 cell line has earned its place as a producer of biotherapeutics. In addition to its ease of growth in serum-free suspension culture and its amenability to transfection, this cell line’s most important attribute is its human origin, which makes it suitable to produce biologics intended for human use. At the present time, the growth and production properties of the HEK293 cell line are inferior to those of non-human cell lines, such as the Chinese hamster ovary (CHO) and the murine myeloma NSO cell lines. However, the modification of genes involved in cellular processes, such as cell proliferation, apoptosis, metabolism, glycosylation, secretion, and protein folding, in addition to bioprocess, media, and vector optimization, have greatly improved the performance of this cell line. This review provides a comprehensive summary of important achievements in HEK293 cell line engineering and on the global engineering approaches and functional genomic tools that have been employed to identify relevant genes for targeted engineering.

scholarly journals MicroRNA-181a Inhibits Activated B-Cell-Like Diffuse Large B-Cell Lymphoma Progression by Repressing CARD11

2019 ◽  
Vol 2019 ◽  
pp. 1-9 ◽  
Author(s):  
Danxia Zhu ◽  
Cheng Fang ◽  
Wenting He ◽  
Chen Wu ◽  
Xiaodong Li ◽  
...  
Keyword(s):  
B Cell ◽  
Binding Site ◽  
Cell Lines ◽  
Expression Vector ◽  
Cell Lymphoma ◽  
B Cell Lymphoma ◽  
Wild Type ◽  
Protein Levels ◽  
Large B Cell Lymphoma ◽  
Large B Cell

We investigated the role of miR-181a in diffuse large B-cell lymphoma (DLBCL) and its potential target genes. miR-181a levels were lower in activated B-cell- (ABC-) like DLBCL cells than that in germinal center B-cell- (GCB-) like DLBCL cells. Overexpression of miR-181a in ABC-like DLBCL cell lines (OCI-LY10 and U2932) resulted in G0/G1 cell cycle arrest, increased apoptosis, and decreased invasiveness. miRNA target prediction programs (miRanda, TargetScan, and miRDB) identified caspase recruitment domain-containing protein 11 (CARD11) as a putative miR-181a target. CARD11 mRNA and protein levels were higher in the ABC-like DLBCL than that in GCB-like DLBCL. Moreover, CARD11 mRNA and protein levels were downregulated in the OCI-LY10 and U2932 cell lines overexpressing miR-181a. Dual luciferase reporter assays confirmed the miR-181a binding site in the CARD11 3′UTR region. OCI-LY10 and U2932 cells transfected with a CARD11 expression vector encoding miR-181a with a mutated binding site showed higher CARD11 protein levels, cell viability, G2/M phase cells, and invasiveness compared to those transfected with a wild-type CARD11 expression vector. Nude mice xenografted with OCI-LY10 cells with overexpressed wild-type miR-181a generated smaller tumors compared to those with overexpressed mutated binding site of CARD11 3′UTR and miR-181a. These results indicate that miR-181a inhibits ABC-like DLBCL by repressing CARD11.

scholarly journals Antibacterial and Cytotoxicity Activities of Major Compounds from Tinospora cordifolia Willd. Growing on Mangifera indica L.

2019 ◽  
Vol 3 (4) ◽  
pp. 32-42
Author(s):  
Thongchai Taechowisan
Keyword(s):  
Antibacterial Activity ◽  
Cell Lines ◽  
Crude Extract ◽  
Hek293 Cell ◽  
Colorimetric Assay ◽  
Mangifera Indica ◽  
Tinospora Cordifolia ◽  
Cytotoxicity Activity ◽  
Stem Extract ◽  
Cytotoxicity Activities

Objective To investigate the major constituents of Tinosporacordifolia Willd. growing on Mangiferaindica, and to evaluate the efficacy of their antibacterial and cytotoxicity activities. Methods The ethanolic stem extract of T. cordifolia was subjected to silica gel 60 column chromatography, thin layer chromatography and medium pressure liquid chromatography for isolation of the major compounds. Identification of purified compounds was achieved by spectroscopic methods.. The crude extract and purified compounds were screened for their antibacterial and cytotoxicity properties using standard procedures. Results Two alkaloids were purified and identified as Magnoflorin (1) and Tembetarine (2). These compounds showed high antibacterial activity against Bacillus cereus and Staphylococcus aureus with both MIC (32-64 µg/ml) and MBC (128-256 µg/ml). The cytotoxicity activity of the purified compounds and crude extract was determined using MTT colorimetric assay against L929 and HEK293 cell lines. This showed weak cytotoxicity activity with IC50 values of 1162.24 to 2290.00 µg/ml and 1376.67 to 2585.06 µg/ml towards L929 and HEK293 cell lines, respectively. Conclusion The major compounds present in ethanolic stem extract of T. cordifolia growing on M. indica were extracted, purified and identified. This study suggests that these compounds exhibit great potential for antibacterial activity with weak cytotoxicity activity. They may be useful for their medicinal functions.

An Eco-friendly Fabrication of Silver Chloride Nanoparticles (AgCLNPs) using Onopordum acanthium L extract Induces Apoptosis in Breast Cancer MDA-MB-232 Cells

2021 ◽  
Author(s):  
Seyedeh Negin Shahcheraghi ◽  
Seyed Ataollah Sadat Shandiz ◽  
Bahareh Pakpour
Keyword(s):  
Breast Cancer ◽  
Real Time ◽  
Cell Lines ◽  
Reverse Transcription ◽  
Hek293 Cell ◽  
Silver Chloride ◽  
Hek293 Cells ◽  
Qrt Pcr ◽  
Reverse Transcription Pcr ◽  
Onopordum Acanthium

Abstract In the current experimental work, silver chloride nanoparticles (AgClNPs) were fabricated using Onopordum acanthium L extract and their apoptotic and cytotoxicity properties on breast cancer MDA_MB232 and normal HEK293 cell lines were also evaluated. AgClNPs formation was determined by X-ray diffraction (XRD), scanning electron microscopy (SEM), and energy dispersive spectroscopy (EDS) profile. Effect of fabricated AgClNPs on MDA_MB232 and HEK293 cells viability was performed using colorimetric MTT assay. Alterations in the mRNA expression levels of CAD and Bax genes in MDA-MB-232 cells were done using quantitative real-time reverse transcription-PCR (qRT-PCR) method. Subsequently, apoptotic properties were determined using flow cytometry and fluorescence microscopy studies. MTT results investigated that AgCLNPs have a significant dose-dependent lethal activity on MDA_MB232 compared to HEK293 cell lines. Quantitative real-time reverse transcription-PCR (qRT-PCR) results have also shown that AgCLNPs could up-regulate the apoptotic Bax and CAD gene expressions in the MDA_MB232 cells. Additionally, apoptotic assessment was performed by cell cycle analysis, annexin V/PI test, Hoescht 33258 dye, acridine orange and ethidium bromide (AO/EB) staining along with the detection of the reactive oxygen species (ROS) generation. Our results suggest that novel silver chloride nanoparticles fabricated by Onopordum acanthium L extract can display some promising cytotoxic properties through inducing apoptosis pathway.

Clonal selection of high producing, stably transfected HEK293 cell lines utilizing modified, high-throughput FACS screening

2011 ◽  
Vol 86 (7) ◽  
pp. 935-941 ◽  
Author(s):  
Michael Song ◽  
Kristin Raphaelli ◽  
Martina L. Jones ◽  
Khosrow Aliabadi-Zadeh ◽  
Kar Man Leung ◽  
...  
Keyword(s):  
Cell Lines ◽  
High Throughput ◽  
Hek293 Cell ◽  
Clonal Selection ◽  
Transfected Hek293 Cell ◽  
Selection Of

c-myb Transactivates the Human Cyclin A1 Promoter and Induces Cyclin A1 Gene Expression

Blood ◽  
1999 ◽  
Vol 94 (12) ◽  
pp. 4255-4262 ◽  
Author(s):  
Carsten Müller ◽  
Rong Yang ◽  
Gregory Idos ◽  
Nicola Tidow ◽  
Sven Diederichs ◽  
...  
Keyword(s):  
Acute Myeloid Leukemia ◽  
Cell Lines ◽  
Myeloid Leukemia ◽  
Expression Vector ◽  
Leukemia Cell ◽  
Myeloid Cell ◽  
Site Directed Mutagenesis ◽  
Cyclin A1 ◽  
High Level ◽  
Acute Myeloid

Cyclin A1 differs from other cyclins in its highly restricted expression pattern. Besides its expression during spermatogenesis, cyclin A1 is also expressed in hematopoietic progenitor cells and in acute myeloid leukemia. We investigated mechanisms that might contribute to cyclin A1 expression in hematopoietic cells. Comparison of cyclin A1 and cyclin A promoter activity in adherent and myeloid leukemia cell lines showed that the cyclin A1 promoter is preferentially active in myeloid cell lines. This preferential activity was present in a small, 335-bp cyclin A1 promoter fragment that contained several potential c-myb binding sites. Coexpression of a c-myb expression vector with the cyclin A1 promoter constructs significantly increased the reporter activity in adherent CV-1 as well as in myeloid U937 cells. Gel-shift assays demonstrated that c-myb could bind to the cyclin A1 promoter at a binding site located near the transcription start site. Site-directed mutagenesis of this site decreased promoter transactivation by 50% in both KCL22 cells that express high levels of c-myb and in CV-1 cells that were transfected with c-myb. In addition, transfection of primary human embryonic fibroblasts with a c-myb expression vector led to induction of the endogenous cyclin A1 gene. Taken together, c-myb can directly transactivate the promoter of cyclin A1, and c-myb might be involved in the high-level expression of cyclin A1 observed in acute myeloid leukemia. These findings suggest that c-myb induces hematopoiesis-specific mechanisms of cell cycle regulation.

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