Protocol to screen for Sorafenib resistance regulators using pooled lentiviral shRNA library and a Sorafenib-resistant hepatocellular carcinoma cell model

Approaches to study therapy resistance in HCC (hepatocellular carcinoma) are limited, especially when using HCC models in vitro. Here, we present a protocol to establish an in vitro Sorafenib-resistant human HCC cell model and conduct an shRNA-mediated synthetic lethal screen in established Sorafeni...

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Detalles Bibliográficos
Autores principales: Gao, Ruize, Tang, Fengyuan, Christofori, Gerhard
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2023
Materias:
Protocol
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10165445/
https://www.ncbi.nlm.nih.gov/pubmed/37126443
http://dx.doi.org/10.1016/j.xpro.2023.102273
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author Gao, Ruize
Tang, Fengyuan
Christofori, Gerhard
author_facet Gao, Ruize
Tang, Fengyuan
Christofori, Gerhard
author_sort Gao, Ruize
collection PubMed
description Approaches to study therapy resistance in HCC (hepatocellular carcinoma) are limited, especially when using HCC models in vitro. Here, we present a protocol to establish an in vitro Sorafenib-resistant human HCC cell model and conduct an shRNA-mediated synthetic lethal screen in established Sorafenib-resistant HCC cell lines to identify critical regulators of Sorafenib resistance. We describe steps for RNA sequencing and functional analysis to reveal the mode of action of potential candidates in conferring therapy resistance to HCC cells. For complete details on the use and execution of this protocol, please refer to Gao et al. (2021a)(1) and Gao et al. (2021b).(2)
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spelling pubmed-101654452023-05-09 Protocol to screen for Sorafenib resistance regulators using pooled lentiviral shRNA library and a Sorafenib-resistant hepatocellular carcinoma cell model Gao, Ruize Tang, Fengyuan Christofori, Gerhard STAR Protoc Protocol Approaches to study therapy resistance in HCC (hepatocellular carcinoma) are limited, especially when using HCC models in vitro. Here, we present a protocol to establish an in vitro Sorafenib-resistant human HCC cell model and conduct an shRNA-mediated synthetic lethal screen in established Sorafenib-resistant HCC cell lines to identify critical regulators of Sorafenib resistance. We describe steps for RNA sequencing and functional analysis to reveal the mode of action of potential candidates in conferring therapy resistance to HCC cells. For complete details on the use and execution of this protocol, please refer to Gao et al. (2021a)(1) and Gao et al. (2021b).(2) Elsevier 2023-04-30 /pmc/articles/PMC10165445/ /pubmed/37126443 http://dx.doi.org/10.1016/j.xpro.2023.102273 Text en © 2023 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
Gao, Ruize
Tang, Fengyuan
Christofori, Gerhard
Protocol to screen for Sorafenib resistance regulators using pooled lentiviral shRNA library and a Sorafenib-resistant hepatocellular carcinoma cell model
title Protocol to screen for Sorafenib resistance regulators using pooled lentiviral shRNA library and a Sorafenib-resistant hepatocellular carcinoma cell model
title_full Protocol to screen for Sorafenib resistance regulators using pooled lentiviral shRNA library and a Sorafenib-resistant hepatocellular carcinoma cell model
title_fullStr Protocol to screen for Sorafenib resistance regulators using pooled lentiviral shRNA library and a Sorafenib-resistant hepatocellular carcinoma cell model
title_full_unstemmed Protocol to screen for Sorafenib resistance regulators using pooled lentiviral shRNA library and a Sorafenib-resistant hepatocellular carcinoma cell model
title_short Protocol to screen for Sorafenib resistance regulators using pooled lentiviral shRNA library and a Sorafenib-resistant hepatocellular carcinoma cell model
title_sort protocol to screen for sorafenib resistance regulators using pooled lentiviral shrna library and a sorafenib-resistant hepatocellular carcinoma cell model
topic Protocol
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10165445/
https://www.ncbi.nlm.nih.gov/pubmed/37126443
http://dx.doi.org/10.1016/j.xpro.2023.102273
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