Journal
FLY
Volume 8, Issue 1, Pages 52-57Publisher
LANDES BIOSCIENCE
DOI: 10.4161/fly.26828
Keywords
RNA-guided DNA cleavage; germline; genomic engineering; engineered endonuclease; CRISPR/Cas9
Categories
Funding
- Mayo Clinic Summer Undergraduate Research Fellowship
- Mayo Graduate School Fellowship
- Mayo Clinic New Investigator Startup Fund
- Richard F. Emslander Career Development Award
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The type II CRISPR/Cas9 system (clustered regularly interspaced short palindromic repeats/CRISPR-associated) has recently emerged as an efficient and simple tool for site-specific engineering of eukaryotic genomes. To improve its applications in Drosophila genome engineering, we simplified the standard two-component CRISPR/Cas9 system by generating a stable transgenic fly line expressing the Cas9 endonuclease in the germline (Vasa-Cas9 line). By injecting vectors expressing engineered target-specific guide RNAs into Vasa-Cas9 fly embryos, mutations were generated from site-specific DNA cleavages and efficiently transmitted into progenies. Because Cas9 endonuclease is the universal component of the type II CRISPR/Cas9 system, site-specific genomic engineering based on this improved platform can be achieved with lower complexity and toxicity, greater consistency, and excellent versatility.
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