Genome engineering of Drosophila with the CRISPR RNA-guided Cas9 nuclease
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...Efficient genome engineering to allow targeted genome modifications in the germ lines of animal models such as fruit flies (113, 114), zebrafish (94, 115), nematodes (116), salamanders (117), and frogs (118, 119) is now possible with the development of the CRISPR-Cas9 technology....
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"Genome engineering of Drosophila wi..." refers background in this paper
...All plasmids used in this study will be made available to academic laboratories through the nonprofit distributor Addgene along with detailed guidance at http://FlyCRISPR.molbio.wisc.edu....
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...…sequence PAM cut site Figure S1 CRISPR RNA/Cas9 molecular reagents used in this study (A) Schematic of the modified CRISPR RNA/Cas9…...
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...Moreover, it may be possible to bias repair to HR over NHEJ by exploiting a Cas9 mutant that induces single-strand DNA breaks (Jinek et al. 2012; Cong et al. 2013; Mali et al. 2013) or conducting Cas9-mediated genome engineering in a genetic Note 1033 background that reduces NHEJ (Beumer et al.…...
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...Recognizing the potential of harnessing this system for precise genome engineering in other organisms, Jinek and colleagues (Jinek et al. 2012) identified a minimal twocomponent system required for the site-specific cleavage of DNA—Cas9 and a chimeric RNA (chiRNA) comprising the crRNA and tracrRNA…...
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...Cleavage also requires that the 39 end of the genomic target sequence contains a 3-basepair (bp) proto-spacer adjacent motif (PAM) sequence, NGG, which differentiates self from invading DNA in the endogenous system (Jinek et al. 2012)....
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"Genome engineering of Drosophila wi..." refers background in this paper
...…findings from mammalian cell culture in which 19- and 118-bp sequences were deleted by cotransfection of Cas9 and two chiRNAs (Cong et al. 2013; Mali et al. 2013) and demonstrate that such “multiplex” genome editing is an effective tool for precisely deleting large genomic regions in organisms....
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...…it may be possible to bias repair to HR over NHEJ by exploiting a Cas9 mutant that induces single-strand DNA breaks (Jinek et al. 2012; Cong et al. 2013; Mali et al. 2013) or conducting Cas9-mediated genome engineering in a genetic Note 1033 background that reduces NHEJ (Beumer et al. 2008)....
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...E-mail: oconnorgiles@wisc.edu Genetics, Vol. 194, 1029–1035 August 2013 1029 eukaryotic genomes, including cultured mammalian cells, human stem cells, and yeast (Cho et al. 2013; Cong et al. 2013; Dicarlo et al. 2013; Ding et al. 2013; Jinek et al. 2013; Mali et al. 2013)....
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"Genome engineering of Drosophila wi..." refers background in this paper
...CRISPRs (clustered regularly interspaced short palindromic repeats) were first identified in Escherichea coli in 1987 and later shown to function as part of an adaptive immune system in bacteria and archaea (Ishino et al. 1987; Makarova et al. 2006; Barrangou et al. 2007)....
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