Endogenous HIF2A reporter systems for high-throughput functional screening.
Zaini, M Nazhif
Patel, Saroor A
Syafruddin, Saiful E
Springer Science and Business Media LLC
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Zaini, M. N., Patel, S. A., Syafruddin, S. E., Rodrigues, P., & Vanharanta, S. (2018). Endogenous HIF2A reporter systems for high-throughput functional screening.. Sci Rep, 8 (1), 12063. https://doi.org/10.1038/s41598-018-30499-2
Tissue-specific transcriptional programs control most biological phenotypes, including disease states such as cancer. However, the molecular details underlying transcriptional specificity is largely unknown, hindering the development of therapeutic approaches. Here, we describe novel experimental reporter systems that allow interrogation of the endogenous expression of HIF2A, a critical driver of renal oncogenesis. Using a focused CRISPR-Cas9 library targeting chromatin regulators, we provide evidence that these reporter systems are compatible with high-throughput screening. Our data also suggests redundancy in the control of cancer type-specific transcriptional traits. Reporter systems such as those described here could facilitate large-scale mechanistic dissection of transcriptional programmes underlying cancer phenotypes, thus paving the way for novel therapeutic approaches.
Cell Line, Tumor, Humans, Carcinoma, Renal Cell, Kidney Neoplasms, Feasibility Studies, Gene Expression Profiling, Transcription, Genetic, Gene Expression Regulation, Neoplastic, Genes, Reporter, Basic Helix-Loop-Helix Transcription Factors, Gene Regulatory Networks, Gene Knock-In Techniques, Genetic Testing, High-Throughput Screening Assays, HEK293 Cells, Carcinogenesis, CRISPR-Cas Systems
Medical Research Council (MC_UU_12022/7)
Majlis Amanah Rakyat (MARA) (unknown)
University of Malaya (UM) (UKM(PER)K019321)
External DOI: https://doi.org/10.1038/s41598-018-30499-2
This record's URL: https://www.repository.cam.ac.uk/handle/1810/279893
Attribution 4.0 International (CC BY 4.0)
Licence URL: https://creativecommons.org/licenses/by/4.0/