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Rapid and precise genome engineering in a naturally short-lived vertebrate

View ORCID ProfileRavi D. Nath, View ORCID ProfileClaire N. Bedbrook, Rahul Nagvekar, Karl Deisseroth, Anne Brunet
doi: https://doi.org/10.1101/2022.05.25.493454
Ravi D. Nath
1Department of Genetics, Stanford University, Stanford, CA 94305, USA
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Claire N. Bedbrook
1Department of Genetics, Stanford University, Stanford, CA 94305, USA
2Department of Bioengineering, Stanford University, Stanford, CA 94305, USA
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Rahul Nagvekar
1Department of Genetics, Stanford University, Stanford, CA 94305, USA
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Karl Deisseroth
2Department of Bioengineering, Stanford University, Stanford, CA 94305, USA
3Department of Psychiatry and Behavioral Sciences, Stanford University, Stanford, CA 94305, USA
4Howard Hughes Medical Institute, Stanford University, Stanford, CA 94305, USA
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Anne Brunet
1Department of Genetics, Stanford University, Stanford, CA 94305, USA
5Glenn Laboratories for the Biology of Aging at Stanford, Stanford, CA 94305, USA
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  • For correspondence: abrunet1@stanford.edu
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Abstract

The African turquoise killifish is a powerful vertebrate system to study complex phenotypes at scale, including aging and age-related disease. Here we develop a rapid and precise CRISPR/Cas9-mediated knock-in approach in the killifish. We show its efficient application to precisely insert fluorescent reporters of different sizes at various genomic loci, to drive cell-type- and tissue-specific expression. This knock-in method should allow the establishment of humanized disease models and the development of cell-type-specific molecular probes for studying complex vertebrate biology.

Competing Interest Statement

The authors have declared no competing interest.

  • Abbreviations

    CN
    cortical nucleus
    CP
    central posterior thalamic nucleus
    Cpost
    posterior commissure
    DIL
    diffuse inferior lobe of hypothalamus
    gl
    glomerular layer
    Ha
    habenular nucleus
    Hc
    caudal hypothalamus
    Hd
    dorsal hypothalamus
    Hv
    ventral hypothalamus
    llf
    lateral longitudinal fascicle
    LR
    lateral recess of diencephalic ventricle
    mlf
    medial longitudinal fascicle
    MO
    medulla oblongata
    NG
    glomerular nucleus
    OB
    olfactory bulb
    ON
    optic nerve
    OT
    optic tectum
    PGZ
    periglomerular gray zone
    Tel
    telencephalon
    Tl
    torus longitudinalis
    TNa
    anterior tuberal nucleus
    TPp
    periventricular nucleus of posterior tuberculum
    Va
    valvula of cerebellum
    VAO
    ventral accessory optic nucleus
  • Copyright 
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    Posted May 26, 2022.
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    Rapid and precise genome engineering in a naturally short-lived vertebrate
    Ravi D. Nath, Claire N. Bedbrook, Rahul Nagvekar, Karl Deisseroth, Anne Brunet
    bioRxiv 2022.05.25.493454; doi: https://doi.org/10.1101/2022.05.25.493454
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    Rapid and precise genome engineering in a naturally short-lived vertebrate
    Ravi D. Nath, Claire N. Bedbrook, Rahul Nagvekar, Karl Deisseroth, Anne Brunet
    bioRxiv 2022.05.25.493454; doi: https://doi.org/10.1101/2022.05.25.493454

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