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

Claire N Bedbrook; Ravi D Nath; Rahul Nagvekar; Karl Deisseroth; Anne Brunet

doi:10.7554/eLife.80639

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

Elife. 2023 May 9:12:e80639. doi: 10.7554/eLife.80639.

Authors

Claire N Bedbrook^#^{1

2}, Ravi D Nath^#¹, Rahul Nagvekar¹, Karl Deisseroth^{2

3

4}, Anne Brunet^{1

5

6}

Affiliations

¹ Department of Genetics, Stanford University, Stanford, United States.
² Department of Bioengineering, Stanford University, Stanford, United States.
³ Department of Psychiatry and Behavioral Sciences, Stanford University, Stanford, United States.
⁴ Howard Hughes Medical Institute, Stanford University, Stanford, United States.
⁵ Glenn Laboratories for the Biology of Aging at Stanford, Stanford, United States.
⁶ Wu Tsai Neurosciences Institute, Stanford University, Stanford, United States.

^# Contributed equally.

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.

Keywords: African killifish; CRISPR/Cas9-mediated knock-in; Nothobranchius furzeri; aging; genetics; genome engineering; genomics.

Publication types

Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't

MeSH terms

Aging / genetics
Animals
Fundulidae*
Genome
Models, Animal
Vertebrates* / genetics

Abstract

Publication types

MeSH terms

Grants and funding