Abstract
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.
Original language | English (US) |
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Pages (from-to) | 52-57 |
Number of pages | 6 |
Journal | Fly |
Volume | 8 |
Issue number | 1 |
DOIs | |
State | Published - 2014 |
Keywords
- CRISPR/Cas9
- Engineered endonuclease
- Genomic engineering
- Germline
- RNA-guided DNA cleavage
ASJC Scopus subject areas
- Insect Science