Cell Reports
Volume 22, Issue 1, 2 January 2018, Pages 232-241
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An Aneuploidy-Free and Structurally Defined Balancer Chromosome Toolkit for Caenorhabditis elegans

https://doi.org/10.1016/j.celrep.2017.12.024Get rights and content
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Highlights

  • Generation of a set of structurally defined and aneuploidy-free balancer chromosomes

  • Reliable and easy analyses of embryonic and early larval lethal phenotypes

  • Utilization of the balancers for isolating lethal alleles using the CRISPR/Cas9 system

Summary

Balancer chromosomes are critical tools for genetic research. In C. elegans, reciprocal translocations that lead to aneuploidy have been widely used to maintain lethal and sterile mutations in stable stocks. Here, we generated a set of aneuploidy-free and structurally defined crossover suppressors that contain two overlapping inversions using the CRISPR/Cas9 system. The toolkit includes 13 crossover suppressors and covers approximately 63% of all C. elegans coding genes. Together with the classical intrachromosomal crossover suppressors, the system now covers 89% of the coding genes. We also labeled the created balancers with fluorescent and phenotypic markers. We show that the crossover suppressors are better for embryonic analysis compared with translocational balancers. Additionally, we demonstrate an efficient method to generate lethal alleles by targeting essential genes on a chromosome balanced with a crossover suppressor. The toolkit will allow more efficient experiments in which lethal and sterile mutants can be analyzed.

Keywords

balancer chromosomes
CRISPR/Cas9
C. elegans
essential gene knockout
chromosomal rearrangements

Cited by (0)

3

Present address: Center for Education in Laboratory Animal Research, Chubu University, Aichi, Japan

4

These authors contributed equally

5

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