Precision genome editing using CRISPR-Cas9 and linear repair templates in C. elegans

Alexandre Paix, Andrew Folkmann, Geraldine Seydoux

Research output: Contribution to journalArticle

Abstract

The ability to introduce targeted edits in the genome of model organisms is revolutionizing the field of genetics. State-of-the-art methods for precision genome editing use RNA-guided endonucleases to create double-strand breaks (DSBs) and DNA templates containing the edits to repair the DSBs. Following this strategy, we have developed a protocol to create precise edits in the C. elegans genome. The protocol takes advantage of two innovations to improve editing efficiency: direct injection of CRISPR-Cas9 ribonucleoprotein complexes and use of linear DNAs with short homology arms as repair templates. The protocol requires no cloning or selection, and can be used to generate base and gene-size edits in just 4 days. Point mutations, insertions, deletions and gene replacements can all be created using the same experimental pipeline.

Original languageEnglish (US)
Pages (from-to)86-93
Number of pages8
JournalMethods
Volume121-122
DOIs
StatePublished - May 15 2017

Keywords

  • C. elegans
  • CRISPR
  • Cas9 RNP
  • Gene conversion
  • Genome editing
  • Homology-dependent repair (HDR)
  • Linear repair templates
  • Short homology

ASJC Scopus subject areas

  • Molecular Biology
  • Biochemistry, Genetics and Molecular Biology(all)

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