Introduction to CRISPR and Its Use in Drosophila
- 1Department of Neuroscience, Brown University, Providence, Rhode Island 02912, USA
- 2Carney Institute for Brain Science, Brown University, Providence, Rhode Island 02912, USA
- 3Pediatrics, Cell & Developmental Biology, University of California, San Diego, La Jolla, California 92093, USA
- 4Biological Sciences, Cell & Developmental Biology, University of California, San Diego, La Jolla, California 92093, USA
- ↵5Correspondence: oconnorgiles{at}brown.edu; jwildonger{at}ucsd.edu
Abstract
The preeminence of Drosophila genetics has led to key discoveries in biology across a variety of fields and disciplines. The advent of CRISPR gene editing has expanded the toolkit of genetic reagents that can be applied to manipulate and observe genes, RNAs, and proteins in an in vivo context. This review describes CRISPR and its use as a transformative gene editing tool in Drosophila. We focus on the canonical pathway in which the Cas9 nuclease is directed to specific sequences by guide RNA (gRNA), where cleavage leads to DNA repair by one of two main cellular pathways: nonhomologous end joining (NHEJ) or homology-directed repair (HDR). The error-prone NHEJ pathway can be appropriated to disrupt targeted sequences, enabling a variety of loss-of-function studies. Induction of the HDR pathway allows precise editing, including defined deletions, the introduction of specific sequence changes, and the incorporation of fluorescent and epitope tags. These approaches have increased the power of Drosophila genetics and been successfully used to conduct in vivo structure–function studies, study disease-associated variants, and follow protein dynamics.
Footnotes
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From the Drosophila Neurobiology collection, edited by Bing Zhang, Ellie Heckscher, Alex C. Keene, and Scott Waddell.
