Efficient Genome Editing in Caenorhabditis elegans with a Toolkit of Dual-Marker Selection Cassettes
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https://doi.org/10.1534/genetics.115.180679Metadata
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Norris, Adam D., Hyun-Min Kim, Mónica P. Colaiácovo, and John A. Calarco. 2015. “Efficient Genome Editing in Caenorhabditis elegans with a Toolkit of Dual-Marker Selection Cassettes.” Genetics 201 (2): 449-458. doi:10.1534/genetics.115.180679. http://dx.doi.org/10.1534/genetics.115.180679.Abstract
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.Other Sources
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4596661/pdf/Terms of Use
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