@article{992936e113874b63acf360c25a651657,
title = "Protocol to screen for Sorafenib resistance regulators using pooled lentiviral shRNA library and a Sorafenib-resistant hepatocellular carcinoma cell model",
abstract = "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",
keywords = "Cancer, Cell Biology, High Throughput Screening, Molecular Biology, RNAseq, Sequencing",
author = "Ruize Gao and Fengyuan Tang and Gerhard Christofori",
note = "Funding Information: We thank A. Kapus, B. Gan, and A. Hergovich for providing plasmid constructs. We thank C. Beisel and the Genomics Facility Basel for RNA and barcode sequencing. We are grateful to Luca Quagliata for sharing the HCC cell lines, David B{\"u}chel for preparing the Sorafenib-resistant cell lines, and Ravi Kiran Reddy Kalathur for the data analysis. This works was supported by the European Research Council (ERC) Synergy Project MERiC and the Swiss National Science Foundation Sinergia Project MERiC. Conceptualization, F.T. R.G. G.C.; Methodology, F.T. R.G.; Investigation, R.G.; Writing – Original Draft, R.G.; Writing – Review & Editing, G.C. F.T. R.G.; Supervision, G.C.; Project Administration, G.C.; Funding Acquisition, G.C. The authors declare no competing interests. Funding Information: We thank A. Kapus, B. Gan, and A. Hergovich for providing plasmid constructs. We thank C. Beisel and the Genomics Facility Basel for RNA and barcode sequencing. We are grateful to Luca Quagliata for sharing the HCC cell lines, David B{\"u}chel for preparing the Sorafenib-resistant cell lines, and Ravi Kiran Reddy Kalathur for the data analysis. This works was supported by the European Research Council (ERC) Synergy Project MERiC and the Swiss National Science Foundation Sinergia Project MERiC . Publisher Copyright: {\textcopyright} 2023 The Author(s)",
year = "2023",
month = jun,
day = "16",
doi = "10.1016/j.xpro.2023.102273",
language = "English",
volume = "4",
journal = "STAR Protocols",
issn = "2666-1667",
publisher = "Cell Press",
number = "2",
}