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  • Title: Highly Efficient, Rapid and Co-CRISPR-Independent Genome Editing in Caenorhabditis elegans.
    Author: Prior H, Jawad AK, MacConnachie L, Beg AA.
    Journal: G3 (Bethesda); 2017 Nov 06; 7(11):3693-3698. PubMed ID: 28893845.
    Abstract:
    We describe a rapid and highly efficient method to generate point mutations in Caenorhabditis elegans using direct injection of CRISPR-Cas9 ribonucleoproteins. This versatile method does not require sensitized genetic backgrounds or co-CRISPR selection-based methods, and represents a single strategy that can be used for creating genomic point mutations, regardless of location. As proof of principle, we show that knock-in mutants more faithfully report variant-associated phenotypes as compared to transgenic overexpression. Data for nine knock-in mutants across five genes are presented that demonstrate high editing efficiencies (60%), a reduced screening workload (24 F1 progeny), and a rapid timescale (4-5 d). This optimized method simplifies genome engineering and is readily adaptable to other model systems.
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