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.
journal_name
Geneticsjournal_title
Geneticsauthors
Norris AD,Kim HM,Colaiácovo MP,Calarco JAdoi
10.1534/genetics.115.180679subject
Has Abstractpub_date
2015-10-01 00:00:00pages
449-58issue
2eissn
0016-6731issn
1943-2631pii
genetics.115.180679journal_volume
201pub_type
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