Generation of a transparent killifish line through multiplex CRISPR/Cas9-mediated gene inactivation
- Leibniz Institute on Aging - Fritz Lipmann Institute, Germany
Abstract
Body pigmentation is a limitation for in vivo imaging and thus for the performance of longitudinal studies in biomedicine. A possibility to circumvent this obstacle is the employment of pigmentation mutants, which are used in fish species like zebrafish and medaka. To address the basis of aging, the short-lived African killifish Nothobranchius furzeri has recently been established as a model organism. Despite its short lifespan, N. furzeri shows typical signs of mammalian aging including telomere shortening, accumulation of senescent cells and loss of regenerative capacity. Here, we report the generation of a transparent N. furzeri line by simultaneous inactivation of three key loci responsible for pigmentation. We demonstrate that this stable line, named klara, can serve as a tool for different applications including behavioral experiments and the establishment of a senescence reporter by integration of a fluorophore into the cdkn1a (p21) locus and in vivo microscopy of the resulting line.
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Article and author information
Author details
Vera L Hopfenmüller
Funding
Carl Zeiss Stiftung (IMPULS,P2019-01-006)
- Christoph Englert
Leibniz Institute on Aging (Graduate Student Fellowship)
- Johannes Krug
The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.
Ethics
Animal experimentation: All fish were maintained in the Nothobranchius facility of the Leibniz Institute on Aging - Fritz Lipmann Institute Jena according to the German Animal Welfare Law. The performed experiments reported here were covered by the animal license FLI-17-016, FLI-20-001 and FLI-20-102, which were approved by the local authorities (Thüringer Landesamt für Verbraucherschutz).
Copyright
© 2023, Krug 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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Generation of a transparent killifish line through multiplex CRISPR/Cas9-mediated gene inactivation
eLife 12:e81549.
https://doi.org/10.7554/eLife.81549
