Distinguishing parallel from series circuits with a double knockdown procedure in human cell lines

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 ovaria...

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Detalles Bibliográficos
Autores principales: Gocher-Demske, Angela M., Dai, Shuhang, Edelman, Arthur M.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2022
Materias:
Protocol
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9719107/
https://www.ncbi.nlm.nih.gov/pubmed/36595936
http://dx.doi.org/10.1016/j.xpro.2022.101890
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author Gocher-Demske, Angela M.
Dai, Shuhang
Edelman, Arthur M.
author_facet Gocher-Demske, Angela M.
Dai, Shuhang
Edelman, Arthur M.
author_sort Gocher-Demske, Angela M.
collection PubMed
description 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)
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spelling pubmed-97191072022-12-04 Distinguishing parallel from series circuits with a double knockdown procedure in human cell lines Gocher-Demske, Angela M. Dai, Shuhang Edelman, Arthur M. STAR Protoc Protocol 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) Elsevier 2022-11-30 /pmc/articles/PMC9719107/ /pubmed/36595936 http://dx.doi.org/10.1016/j.xpro.2022.101890 Text en © 2022 The Author(s) https://creativecommons.org/licenses/by-nc-nd/4.0/This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
spellingShingle Protocol
Gocher-Demske, Angela M.
Dai, Shuhang
Edelman, Arthur M.
Distinguishing parallel from series circuits with a double knockdown procedure in human cell lines
title Distinguishing parallel from series circuits with a double knockdown procedure in human cell lines
title_full Distinguishing parallel from series circuits with a double knockdown procedure in human cell lines
title_fullStr Distinguishing parallel from series circuits with a double knockdown procedure in human cell lines
title_full_unstemmed Distinguishing parallel from series circuits with a double knockdown procedure in human cell lines
title_short Distinguishing parallel from series circuits with a double knockdown procedure in human cell lines
title_sort distinguishing parallel from series circuits with a double knockdown procedure in human cell lines
topic Protocol
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9719107/
https://www.ncbi.nlm.nih.gov/pubmed/36595936
http://dx.doi.org/10.1016/j.xpro.2022.101890
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