Abstract
Perturbation of genes in cultured cells is useful for gene function discovery, combinatorial screening, and other applications. In contrast to RNAi the CRISPR system targets genomic sequence. CRISPR can be used to make double strand breaks in genomic DNA to induce mutations in specific gene targets, including knockout mutations or more specific gene engineering (knock-in). Using nuclease-dead forms of Cas9 (dCas9), the CRISPR system can also be used to down- or up-regulate transcription of specific targets. A comparison among RNAi and various CRISPR methods is presented, along with practical information regarding plasmid vectors and protocols relevant to use of the CRISPR system and detection of mutations in Drosophila cultured cells.
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