Ingestion of single guide RNAs induces gene overexpression and extends lifespan in Caenorhabditis elegans via CRISPR activation
Abstract
Inhibition of gene expression in Caenorhabditis elegans, a versatile model organism for studying the genetics of development and aging, is achievable by feeding nematodes with bacteria expressing specific dsRNAs. Overexpression of hypoxia-inducible factor 1 (hif-1) or heat-shock factor 1 (hsf-1) by conventional transgenesis has previously been shown to promote nematodal longevity. However, it is unclear whether other methods of gene overexpression are feasible, particularly with the advent of CRISPR-based techniques. Here, we show that feeding C. elegans engineered to stably express a Cas9-derived synthetic transcription factor with bacteria expressing promoter-specific single guide RNAs (sgRNAs) also allows activation of gene expression. We demonstrate that CRISPR activation via ingested sgRNAs specific for the respective promoter regions of hif-1 or hsf-1 increases gene expression and extends lifespan of C. elegans. Furthermore, and as an in silico resource for future studies aiming to use CRISPR activation in C. elegans, we provide predicted promoter-specific sgRNA target sequences for >13,000 C. elegans genes with experimentally defined transcription start sites. We anticipate that the approach and components described herein will help to facilitate genome-wide gene overexpression studies, for example, to identify modulators of aging or other phenotypes of interest, by enabling induction of transcription by feeding of sgRNA-expressing bacteria to nematodes. Show more
Permanent link
https://doi.org/10.3929/ethz-b-000555534Publication status
publishedExternal links
Journal / series
Journal of Biological ChemistryVolume
Pages / Article No.
Publisher
American Society for Biochemistry and Molecular BiologySubject
aging; C. elegans; CRISPR/Cas9; gene expression; synthetic biology; HIF-1; HSF-1Organisational unit
03976 - Ristow, Michael (ehemalig) / Ristow, Michael (former)
09598 - Ewald, Collin Y. (ehemalig) / Ewald, Collin Y. (former)
Funding
156031 - Molekulare Charakterisierung neuer Spezies-unabhängiger Regulatoren systemischer Alterung (SNF)
204511 - Transcriptional regulators of systemic aging (SNF)
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