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Efficient CRISPR/Cas9 plasmids for rapid and versatile genome editing in Drosophila.

ABSTRACT: The CRISPR-associated RNA-guided nuclease Cas9 has emerged as a powerful tool for genome engineering in a variety of organisms. To achieve efficient gene targeting rates in Drosophila, current approaches require either injection of in vitro transcribed RNAs or injection into transgenic Cas9-expressing embryos. We report a simple and versatile alternative method for CRISPR-mediated genome editing in Drosophila using bicistronic Cas9/sgRNA expression vectors. Gene targeting with this single-plasmid injection approach is as efficient as in transgenic nanos-Cas9 embryos and allows the isolation of targeted knock-out and knock-in alleles by molecular screening within 2 months. Our strategy is independent of genetic background and does not require prior establishment of transgenic flies.

SUBMITTER: Gokcezade J

PROVIDER: S-EPMC4232553 | biostudies-literature | 2014 Sep

REPOSITORIES: biostudies-literature

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Efficient CRISPR/Cas9 plasmids for rapid and versatile genome editing in Drosophila.

Gokcezade Joseph J Sienski Grzegorz G Duchek Peter P

G3 (Bethesda, Md.) 20140917 11

The CRISPR-associated RNA-guided nuclease Cas9 has emerged as a powerful tool for genome engineering in a variety of organisms. To achieve efficient gene targeting rates in Drosophila, current approaches require either injection of in vitro transcribed RNAs or injection into transgenic Cas9-expressing embryos. We report a simple and versatile alternative method for CRISPR-mediated genome editing in Drosophila using bicistronic Cas9/sgRNA expression vectors. Gene targeting with this single-plasmi ...[more]

PMID: 25236734

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