Efficient CRISPR-Cas9 editing of major evolutionary loci in sticklebacks.

ABSTRACT:

Background

Stickleback fish are widely used to study the genetic and ecological basis of phenotypic evolution. Although several major loci have now been identified that contribute to evolutionary differences between wild populations, further study of the phenotypes associated with particular genes and mutations has been limited by the difficulty of generating targeted mutations at precise locations in the stickleback genome.

Approach and aims

We compared different methods of expressing single-guide RNAs (sgRNAs) and Cas9 activity in fertilized stickleback eggs. We used an easily scored pigmentation gene (SLC24A5) to screen for molecular lesions, phenotypic effects, and possible germline transmission of newly induced alleles. We then used the optimized CRISPR methods t

SUBMITTER: Wucherpfennig JI 

PROVIDER: S-EPMC8664273 | biostudies-literature | 2019 Jan

REPOSITORIES: biostudies-literature