title: Novel synthetic lethality screening method identifies TIP60-dependent radiation sensitivity in the absence of BAF180.
abstract: In recent years, research into synthetic lethality and how it can be exploited in cancer treatments has emerged as major focus in cancer research. However, the lack of a simple to use, sensitive and standardised assay to test for synthetic interactions has been slowing the efforts. Here we present a novel approach to synthetic lethality screening based on co-culturing two syngeneic cell lines containing individual fluorescent tags. By associating shRNAs for a target gene or control to individual fluorescence labels, we can easily follow individual cell fates upon siRNA treatment and high content imaging. We have demonstrated that the system can recapitulate the functional defects of the target gene depletion and is capable of discovering novel synthetic interactors and phenotypes. In a trial screen, we show that TIP60 exhibits synthetic lethality interaction with BAF180, and that in the absence of TIP60, there is an increase micronuclei dependent on the level of BAF180 loss, significantly above levels seen with BAF180 present. Moreover, the severity of the interactions correlates with proxy measurements of BAF180 knockdown efficacy, which may expand its usefulness to addressing synthetic interactions through titratable hypomorphic gene expression.
authors.affiliation: Genome Damage and Stability Centre, School of Life Sciences, University of Sussex, Brighton BN1 RQ9, United Kingdom.
authors.forename: Suzanna R
authors.initials: SR
authors.lastname: Hopkins
authors.affiliation: Genome Damage and Stability Centre, School of Life Sciences, University of Sussex, Brighton BN1 RQ9, United Kingdom.
authors.forename: Grant A
authors.initials: GA
authors.lastname: McGregor
authors.affiliation: Genome Damage and Stability Centre, School of Life Sciences, University of Sussex, Brighton BN1 RQ9, United Kingdom.
authors.forename: Johanne M
authors.initials: JM
authors.lastname: Murray
authors.affiliation: Genome Damage and Stability Centre, School of Life Sciences, University of Sussex, Brighton BN1 RQ9, United Kingdom.
authors.forename: Jessica A
authors.initials: JA
authors.lastname: Downs
authors.affiliation: Genome Damage and Stability Centre, School of Life Sciences, University of Sussex, Brighton BN1 RQ9, United Kingdom; Department of Clinical and Experimental Medicine, Brighton and Sussex Medical School, University of Sussex, Brighton BN1 9PX, United Kingdom. Electronic address: V.Savic@bsms.ac.uk.
authors.forename: Velibor
authors.initials: V
authors.lastname: Savic
mesh_terms: D045744:Cell Line, Tumor; D002470:Cell Survival / Q000528:radiation effects; D004268:DNA-Binding Proteins; D005456:Fluorescent Dyes / Q000378:metabolism; D015870:Gene Expression; D057166:High-Throughput Screening Assays; D051548:Histone Acetyltransferases / Q000037:antagonists & inhibitors / Q000235:genetics* / Q000378:metabolism; D006801:Humans; D000074443:Lysine Acetyltransferase 5; D015162:Micronucleus Tests; D057054:Molecular Imaging; D009687:Nuclear Proteins / Q000172:deficiency / Q000235:genetics*; D010006:Osteoblasts / Q000378:metabolism / Q000473:pathology / Q000528:radiation effects*; D034741:RNA, Small Interfering / Q000235:genetics / Q000378:metabolism; D011836:Radiation Tolerance; D013194:Staining and Labeling / Q000379:methods; D000072020:Synthetic Lethal Mutations; D014157:Transcription Factors / Q000172:deficiency / Q000235:genetics*
chemical_list: D004268:DNA-Binding Proteins; D005456:Fluorescent Dyes; D009687:Nuclear Proteins; C102195:PBRM1 protein, human; D034741:RNA, Small Interfering; D014157:Transcription Factors; D051548:Histone Acetyltransferases; C098489:KAT5 protein, human; D000074443:Lysine Acetyltransferase 5