CRISPR-mediated genetic interaction profiling identifies RNA binding proteins controlling metazoan fitness
- Harvard University, United States
- University of Toronto, Canada
Abstract
Genetic interaction screens have aided our understanding of complex genetic traits, diseases, and biological pathways. However, approaches for synthetic genetic analysis with null-alleles in metazoans have not been feasible. Here, we present a CRISPR/Cas9-based Synthetic Genetic Interaction (CRISPR-SGI) approach enabling systematic double-mutant generation. Applying this technique in Caenorhabditis elegans, we comprehensively screened interactions within a set of 14 conserved RNA binding protein genes, generating all possible single and double mutants. Many double mutants displayed fitness defects, revealing synthetic interactions. For one interaction between the MBNL1/2 ortholog mbl-1 and the ELAVL ortholog exc-7, double mutants displayed a severely shortened lifespan. Both genes are required for regulating hundreds of transcripts and isoforms, and both may play a critical role in lifespan extension through insulin signaling. Thus, CRISPR-SGI reveals a rich genetic interaction landscape between RNA binding proteins in maintaining organismal health, and will serve as a paradigm applicable to other biological questions.
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Author details
Funding
NIH Office of the Director (NIH Early Independence Award DP5OD009153)
- John Calarco
Harvard University (Bauer Fellows Program)
- John Calarco
University of Toronto
- John Calarco
Charles King postdoctoral fellowship
- Adam D Norris
NSERC (Discovery Grant RGPIN-2017-06573)
- John Calarco
The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.
Copyright
© 2017, Norris et al.
This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.
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CRISPR-mediated genetic interaction profiling identifies RNA binding proteins controlling metazoan fitness
eLife 6:e28129.
https://doi.org/10.7554/eLife.28129
