October 2020 Newsletter

"One to Watch" in the
Nature Spinoff Prize 2020

USE OF RJH TRANSFECTION REAGENTS IN siRNA LIBRARY SCREENS

Having the ability to screen large numbers of compounds in one attempt enables head-to-head comparisons in an expeditious manner. One has to rely on a reproducible procedure and delivery that are similar for each member of the library component. This is especially the case when transfection reagents are used and robust performance is expected for the delivery of each library member. Work with large libraries also poses a cost burden that has to be minimized for long term utility of such screens.

The RJH transfection reagents are particularly suitable for screening nucleic acid libraries. They are provided in homogenous solutions that minimize well-to-well variations during screens. They can be formulated to function in different formats, either delivering library members on their own or in combination with other reagents for synergistic activities. They are also cost-effective to reduce the overall costs of the screens.

Library Screen in Normal Fibroblasts and Transformed Cells

siRNA delivered by in vivo RNA-Fect (library screen) — Relative growth of MDA-MB-435 cells and skin fibroblasts after treatment with apoptosis-related siRNAs delivered with In Vivo RNA-Fect. The cell growth was normalized against non-treated cells (taken as 100%). The siRNA library was from Dharmacon (siGENOME Human Apoptosis siRNA Library). While fibroblast cells did not show major responses to siRNA treatment, transformed cells displayed lower growth with numerous siRNA treatments.

Library Screen in Leukemic Cells

siRNA delivery using Leu-Fect A for leukemia cells library screen — Relative growth of leukemic RS4;11 cells after treatment with apoptosis-related siRNAs delivered with Leu-Fect A. The cell growth was normalized against non-treated cells (taken as 100%). Treatment results with the positive control treatment doxorubicin are in red circles. Most likely positive hits are shown in the shaded box.

Library Screen in Breast Cancer Cells in Combination with TRAIL Protein

apoptosis siRNA and TRAIL protein treatment of breast cancer cells — Relative growth of MDA-MB-231 cells after treatment with a combination of TRAIL protein and apoptosis-related siRNAs. The cell growth was normalized against non-treated cells (taken as 100%). The siRNA library was from Dharmacon (siGENOME Human Apoptosis siRNA Library). Response with TRAIL protein alone is shown in red dots.

CONSIDERATIONS FOR OPTIMIZING NON-VIRAL TRANSFECTIONS IN CULTURE

The efficiency of all transfection reagents depends on cell type to be modified, nature of polynucleotide to be delivered, and cell culture conditions. The transfection conditions for each cell type should be optimized to ensure maximal gene expression (or silencing) while minimizing cellular toxicity. We have complied a list of factors that should be considered for optimum transfection, which can be accessed here.

Take a look at 'Electrospun gelatine matrices with bioactive pDNA polyplexes', a 2020 publication highlighting RJH Product "All-Fect" for electrospun matrices suitable for implantation.

All-Fect transfection reagent RJH Biosciences

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