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

Author:

Bedbrook Claire N12ORCID,Nath Ravi D1ORCID,Nagvekar Rahul1ORCID,Deisseroth Karl234,Brunet Anne156ORCID

Affiliation:

1. Department of Genetics, Stanford University

2. Department of Bioengineering, Stanford University

3. Department of Psychiatry and Behavioral Sciences, Stanford University

4. Howard Hughes Medical Institute, Stanford University

5. Glenn Laboratories for the Biology of Aging at Stanford

6. Wu Tsai Neurosciences Institute, Stanford University

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 in order 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.

Funder

National Institutes of Health

Glenn Foundation for Medical Research

Simons Foundation

Chan Zuckerberg Biohub - San Francisco

Knight-Hennessy Scholars Graduate Fellowship

Helen Hay Whitney Foundation

Wu Tsai Neurosciences Institute Interdisciplinary Scholar Award

Iqbal Farrukh & Asad Jamal Center for Cognitive Health in Aging

Knight Initiative for Brain Resilience Scholar Award

Publisher

eLife Sciences Publications, Ltd

Subject

General Immunology and Microbiology,General Biochemistry, Genetics and Molecular Biology,General Medicine,General Neuroscience

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