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EP20169
Abstract: Successful RNAi experiments and large-scale siRNA screens require efficient delivery of highly functional and specific nucleic acids including siRNA oligonucleotides, shRNA vectors, or micro RNAs into an appropriate cell system. Cell types relevant for immunological research, such as primary T cells and several suspension cell lines, are poorly accessible using reagent-based transfection approaches.
Nucleofection® is an established method for the effective, non-viral transfection of nucleic acids into difficult-to-transfect cell types including primary cells. With the expansion of the technology to a 96-well format (Fig. 1) highthroughput applications, such as siRNA or shRNA library screenings can now be performed in these cell types rendering target validation and identification possible in cell types highly relevant for medical research. Integration of the 96-well Shuttle® into a liquid handling workstation allows for a fully automated screening approach.
Using the powerful combination of highly functional Dharmacon siGENOME® siRNA reagents and 96-well nucleofection®, we here present data showing the efficient siRNA-mediated gene knockdown in various cell lines and primary cells.
Our studies focus on Jurkat T-cells which are derived from a human acute T-cell leukemia and are extensively used in the study of T-cell signaling and cancer drug development.
Summary: Successful RNAi experiments and large-scale siRNA screens require efficient delivery of highly functional and specific nucleic acids including siRNA oligonucleotides, shRNA vectors, or micro RNAs into an appropriate cell system. Cell types relevant for immunological research, such as primary T cells and several suspension cell lines, are poorly accessible using reagent-based transfection approaches.
Nucleofection® is an established method for the effective, non-viral transfection of nucleic acids into difficult-to-transfect cell types including primary cells. With the expansion of the technology to a 96-well format (Fig. 1) highthroughput applications, such as siRNA or shRNA library screenings can now be performed in these cell types rendering target validation and identification possible in cell types highly relevant for medical research. Integration of the 96-well Shuttle® into a liquid handling workstation allows for a fully automated screening approach.
Using the powerful combination of highly functional Dharmacon siGENOME® siRNA reagents and 96-well nucleofection®, we here present data showing the efficient siRNA-mediated gene knockdown in various cell lines and primary cells.
Our studies focus on Jurkat T-cells which are derived from a human acute T-cell leukemia and are extensively used in the study of T-cell signaling and cancer drug development.
Summary: Successful RNAi experiments and large-scale siRNA screens require efficient delivery of highly functional and specific nucleic acids including siRNA oligonucleotides, shRNA vectors, or micro RNAs into an appropriate cell system. Cell types relevant for immunological research, such as primary T cells and several suspension cell lines, are poorly accessible using reagent-based transfection approaches.
This work is licensed under a Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International License.
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