title: Cas9-Mediated Genome Engineering in Drosophila melanogaster.
abstract: The recent development of the CRISPR-Cas9 system for genome engineering has revolutionized our ability to modify the endogenous DNA sequence of many organisms, including Drosophila This system allows alteration of DNA sequences in situ with single base-pair precision and is now being used for a wide variety of applications. To use the CRISPR system effectively, various design parameters must be considered, including single guide RNA target site selection and identification of successful editing events. Here, we review recent advances in CRISPR methodology in Drosophila and introduce protocols for some of the more difficult aspects of CRISPR implementation: designing and generating CRISPR reagents and detecting indel mutations by high-resolution melt analysis.
authors: Department of Genetics, Harvard Medical School, Boston, Massachusetts 02115;
authors: Benjamin E
authors: BE
authors: Housden
authors: Department of Genetics, Harvard Medical School, Boston, Massachusetts 02115; Howard Hughes Medical Institute, Harvard Medical School, Boston, Massachusetts 02115.
authors: Norbert
authors: N
authors: Perrimon
mesh_terms: D000818:Animals; D064113:CRISPR-Cas Systems; D004331:Drosophila melanogaster / Q000235:genetics*; D000072669:Gene Editing / Q000379:methods*; D011995:Recombination, Genetic