CRISPR-mediated genetic interaction profiling identifies RNA binding proteins controlling metazoan fitness.
Elife
; 62017 07 18.
Article
in En
| MEDLINE
| ID: mdl-28718764
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.
Key words
Full text:
1
Collection:
01-internacional
Database:
MEDLINE
Main subject:
RNA-Binding Proteins
/
Caenorhabditis elegans
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Caenorhabditis elegans Proteins
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RNA Interference
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High-Throughput Screening Assays
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Clustered Regularly Interspaced Short Palindromic Repeats
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Mutation
Limits:
Animals
Language:
En
Journal:
Elife
Year:
2017
Document type:
Article
Affiliation country:
United States