Cas9-based genome editing in Drosophila.
Our ability to modify the Drosophila genome has recently been revolutionized by the development of the CRISPR system. The simplicity and high efficiency of this system allows its widespread use for many different applications, greatly increasing the range of genome modification experiments that can be performed. Here, we first discuss some general design principles for genome engineering experiments in Drosophila and then present detailed protocols for the production of CRISPR reagents and screening strategies to detect successful genome modification events in both tissue culture cells and animals.
Department of Genetics, Harvard Medical School, Boston, Massachusetts, USA. Electronic address: bhousden@genetics.med.harvard.edu.
Benjamin E
BE
Housden
Department of Genetics, Harvard Medical School, Boston, Massachusetts, USA.
Shuailiang
S
Lin
Department of Genetics, Harvard Medical School, Boston, Massachusetts, USA; Howard Hughes Medical Institute, Harvard Medical School, Boston, Massachusetts, USA.
Norbert
N
Perrimon
D000818:Animals; D064113:CRISPR-Cas Systems; D003001:Cloning, Molecular / Q000379:methods; D064112:Clustered Regularly Interspaced Short Palindromic Repeats; D004330:Drosophila / Q000235:genetics*; D018390:Gene Targeting / Q000379:methods*; D005818:Genetic Engineering / Q000379:methods*; D005822:Genetic Vectors / Q000235:genetics; D016678:Genome; D059765:Homologous Recombination; D016296:Mutagenesis; D000094704:RNA, Guide, CRISPR-Cas Systems / Q000235:genetics
D000094704:RNA, Guide, CRISPR-Cas Systems
CRISPR; Drosophila; Genome editing; Genome engineering; High-resolution melt assays; Homologous recombination; Methods