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Efficient Genome Editing in Caenorhabditis elegans with a Toolkit of Dual-Marker Selection Cassettes.

Publication ,  Journal Article
Norris, AD; Kim, H-M; Colaiácovo, MP; Calarco, JA
Published in: Genetics
October 2015

Use of the CRISPR/Cas9 RNA-guided endonuclease complex has recently enabled the generation of double-strand breaks virtually anywhere in the C. elegans genome. Here, we present an improved strategy that makes all steps in the genome editing process more efficient. We have created a toolkit of template-mediated repair cassettes that contain an antibiotic resistance gene to select for worms carrying the repair template and a fluorescent visual marker that facilitates identification of bona fide recombinant animals. Homozygous animals can be identified as early as 4-5 days post-injection, and minimal genotyping by PCR is required. We demonstrate that our toolkit of dual-marker vectors can generate targeted disruptions, deletions, and endogenous tagging with fluorescent proteins and epitopes. This strategy should be useful for a wide variety of additional applications and will provide researchers with increased flexibility when designing genome editing experiments.

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Published In

Genetics

DOI

EISSN

1943-2631

ISSN

0016-6731

Publication Date

October 2015

Volume

201

Issue

2

Start / End Page

449 / 458

Related Subject Headings

  • Recombinational DNA Repair
  • RNA Editing
  • Homologous Recombination
  • Genotype
  • Genome
  • Gene Targeting
  • Developmental Biology
  • DNA Breaks, Double-Stranded
  • Caenorhabditis elegans
  • CRISPR-Cas Systems
 

Citation

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Norris, A. D., Kim, H.-M., Colaiácovo, M. P., & Calarco, J. A. (2015). Efficient Genome Editing in Caenorhabditis elegans with a Toolkit of Dual-Marker Selection Cassettes. Genetics, 201(2), 449–458. https://doi.org/10.1534/genetics.115.180679
Norris, Adam D., Hyun-Min Kim, Mónica P. Colaiácovo, and John A. Calarco. “Efficient Genome Editing in Caenorhabditis elegans with a Toolkit of Dual-Marker Selection Cassettes.Genetics 201, no. 2 (October 2015): 449–58. https://doi.org/10.1534/genetics.115.180679.
Norris AD, Kim H-M, Colaiácovo MP, Calarco JA. Efficient Genome Editing in Caenorhabditis elegans with a Toolkit of Dual-Marker Selection Cassettes. Genetics. 2015 Oct;201(2):449–58.
Norris, Adam D., et al. “Efficient Genome Editing in Caenorhabditis elegans with a Toolkit of Dual-Marker Selection Cassettes.Genetics, vol. 201, no. 2, Oct. 2015, pp. 449–58. Epmc, doi:10.1534/genetics.115.180679.
Norris AD, Kim H-M, Colaiácovo MP, Calarco JA. Efficient Genome Editing in Caenorhabditis elegans with a Toolkit of Dual-Marker Selection Cassettes. Genetics. 2015 Oct;201(2):449–458.

Published In

Genetics

DOI

EISSN

1943-2631

ISSN

0016-6731

Publication Date

October 2015

Volume

201

Issue

2

Start / End Page

449 / 458

Related Subject Headings

  • Recombinational DNA Repair
  • RNA Editing
  • Homologous Recombination
  • Genotype
  • Genome
  • Gene Targeting
  • Developmental Biology
  • DNA Breaks, Double-Stranded
  • Caenorhabditis elegans
  • CRISPR-Cas Systems