Distinguishing parallel from series circuits with a double knockdown procedure in human cell lines
Related Experiment Video
Updated: Aug 15, 2025
Rapid Development of Cell State Identification Circuits with Poly-Transfection
Published on: February 24, 2023
Angela M Gocher-Demske1, Shuhang Dai2, Arthur M Edelman2
1Department of Immunology, University of Pittsburgh School of Medicine, Pittsburgh, PA 15213, USA; Tumor Microenvironment Center, University of Pittsburgh Medical Center (UPMC) Hillman Cancer Center, Pittsburgh, PA 15232, USA.
Abstract:
Signaling cascades can act in series or in parallel. Here, we describe a convenient and robust protocol for dual, sequential knockdown of two proteins using RNA interference. We detail the steps for a quantitative mapping of signaling circuitry. We used this approach to study kinases in human ovarian cancer cells, but the protocol can be applied to many other posttranslational modifications. For complete details on the use and execution of this protocol, please refer to Gocher et al. (2017).1.
More Related Videos
13:54Preparation of Cell-lines for Conditional Knockdown of Gene Expression and Measurement of the Knockdown Effects on E4orf4-Induced Cell Death
Published on: October 21, 2012
10:45Double In Utero Electroporation to Target Temporally and Spatially Separated Cell Populations
Published on: June 14, 2020
