Abstract
A universal process in experimental biology is the use of engineered cells; more often, stably or transiently transfected cells are generated for the purpose. Therefore, it is important that cell health assessment is conducted to check for stress mediated by induction of heat shock proteins (Hsps). For this purpose, we have developed an integrated platform that would enable a direct assessment of transfection efficiency (TE) combined with cellular toxicity and stress response. We make use of automated microscopy and high content analysis to extract from the same well a multiplexed readout to assess and determine optimal chemical transfection conditions. As a proof of concept, we investigated seven commercial reagents, in a matrix of dose and time, to study transfection of an EGFP DNA plasmid into HeLa cells and their consequences on health and fitness; where we scored for cellular proliferation, EGFP positive cells, and induction of Hsp10 and Hsp70 as makers of stress responses. FuGENE HD emerged as the most optimal reagent with no apparent side effects suitable for performing microtiter based miniaturized transfection for both chemical and RNAi screening. In summary, we report on a high content assay method to assess cellular overall fitness upon chemical transfection.
Keywords: Chemical transfection, HCA, HCS, Hsp10, Hsp70, cell stress, INCA2000, INCA6000.
Call for Papers in Thematic Issues
Advances in the design of antibody & protein with conformational dynamics and artificial intelligence approaches
“Antibodies & Protein Design” section focuses on the utilization of multiple strategies to engineer and optimize antibodies and proteins that serve diverse analytical strategies, such as combinatorial protein design, structure-based design, sequence-based design, and other techniques that incorporate principles of protein-protein interactions, allosteric regulation, and post-translational modifications. Example applications include ...read more
Artificial Intelligence Methods for Biomedical, Biochemical and Bioinformatics Problems
Recently, a large number of technologies based on artificial intelligence have been developed and applied to solve a diverse range of problems in the areas of biomedical, biochemical and bioinformatics problems. By utilizing powerful computing resources and massive amounts of data, methods based on artificial intelligence can significantly improve the ...read more
Exploring Spectral Graph Theory in Combinatorial Chemistry
Combinatorial chemistry involves the synthesis and analysis of a large number of diverse compounds simultaneously. Traditional methods rely on brute-force experimentation, which can be time-consuming and resource-intensive. Spectral graph theory, a branch of mathematics dealing with the properties of graphs in relation to the eigenvalues and eigenvectors of matrices associated ...read more
Intersection of Chemical Graph Theory and Molecular Sciences
Chemical graph theory has become increasingly critical in molecular sciences, offering advanced capabilities in the modeling, design, and analysis of chemical structures via molecular graphs. This special issue aims to not only explore these fundamental aspects but also to push the boundaries of CCHTS applications. By integrating the concept of ...read more
Related Journals
Anti-Cancer Agents in Medicinal Chemistry
Anti-Inflammatory & Anti-Allergy Agents in Medicinal Chemistry
Current Analytical Chemistry
Current Computer-Aided Drug Design
Current Bioactive Compounds
Current Cancer Drug Targets
Current Cancer Therapy Reviews
Current Diabetes Reviews
Current Drug Safety
Current Drug Targets
Related Books
Organic Spectroscopy Technology and Applications
Andrographolide and its Analogs: Botanical Sources, Phytochemistry, Pharmacology, and Biotechnology
Metabolites of Medicinal Plants: Insightful Approaches
2-Deoxy-D-Glucose: Chemistry and Biology
Chiral Ionic Liquids: Applications in Chemistry and Technology
Anthocyanins: Pharmacology and Nutraceutical Importance
Therapeutic Insights into Herbal Medicine through the Use of Phytomolecules
The Roles and Responsibilities of Clinical Pharmacists in Hospital Settings
Bioactive Compounds from Medicinal Plants for Cancer Therapy and Chemoprevention
Biochar - Solid Carbon for Sustainable Agriculture
36
