Novel synthetic lethality screening method identifies TIP60-dependent radiation sensitivity in the absence of BAF180

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...

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Bibliographic Details
Published in:DNA repair Vol. 46; pp. 47 - 54
Main Authors: Hopkins, Suzanna R., McGregor, Grant A., Murray, Johanne M., Downs, Jessica A., Savic, Velibor
Format: Journal Article
Language:English
Published: Netherlands Elsevier B.V 01.10.2016
Subjects:
Cell Line, Tumor
Cell Survival - radiation effects
DNA-Binding Proteins
Fluorescent Dyes - metabolism
Gene Expression
High-Throughput Screening Assays
Histone Acetyltransferases - antagonists & inhibitors
Histone Acetyltransferases - genetics
Histone Acetyltransferases - metabolism
Humans
Lysine Acetyltransferase 5
Micronucleus Tests
Molecular Imaging
Nuclear Proteins - deficiency
Nuclear Proteins - genetics
Osteoblasts - metabolism
Osteoblasts - pathology
Osteoblasts - radiation effects
Radiation Tolerance
RNA, Small Interfering - genetics
RNA, Small Interfering - metabolism
Staining and Labeling - methods
Synthetic Lethal Mutations
Transcription Factors - deficiency
Transcription Factors - genetics
ISSN:1568-7864, 1568-7856
Online Access:Get full text
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Summary: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.
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ISSN:1568-7864
1568-7856
DOI:10.1016/j.dnarep.2016.05.030