title: CRISPR/Cas9 mediates efficient conditional mutagenesis in Drosophila.
abstract: Existing transgenic RNA interference (RNAi) methods greatly facilitate functional genome studies via controlled silencing of targeted mRNA in Drosophila. Although the RNAi approach is extremely powerful, concerns still linger about its low efficiency. Here, we developed a CRISPR/Cas9-mediated conditional mutagenesis system by combining tissue-specific expression of Cas9 driven by the Gal4/upstream activating site system with various ubiquitously expressed guide RNA transgenes to effectively inactivate gene expression in a temporally and spatially controlled manner. Furthermore, by including multiple guide RNAs in a transgenic vector to target a single gene, we achieved a high degree of gene mutagenesis in specific tissues. The CRISPR/Cas9-mediated conditional mutagenesis system provides a simple and effective tool for gene function analysis, and complements the existing RNAi approach.
authors.affiliation: School of Life Sciences, Tsinghua University, Beijing 100084, China.
authors.forename: Zhaoyu
authors.initials: Z
authors.lastname: Xue
authors.affiliation: School of Life Sciences, Tsinghua University, Beijing 100084, China.
authors.forename: Menghua
authors.initials: M
authors.lastname: Wu
authors.affiliation: School of Life Sciences, Tsinghua University, Beijing 100084, China.
authors.forename: Kejia
authors.initials: K
authors.lastname: Wen
authors.affiliation: School of Life Sciences, Tsinghua University, Beijing 100084, China.
authors.forename: Menda
authors.initials: M
authors.lastname: Ren
authors.affiliation: School of Life Sciences, Tsinghua University, Beijing 100084, China.
authors.forename: Li
authors.initials: L
authors.lastname: Long
authors.affiliation: School of Life Sciences, Tsinghua University, Beijing 100084, China.
authors.forename: Xuedi
authors.initials: X
authors.lastname: Zhang
authors.affiliation: School of Life Sciences, Tsinghua University, Beijing 100084, China gaogu@mail.tsinghua.edu.cn.
authors.forename: Guanjun
authors.initials: G
authors.lastname: Gao
mesh_terms: D000818:Animals; D064113:CRISPR-Cas Systems; D004330:Drosophila / Q000235:genetics*; D018390:Gene Targeting / Q000379:methods*; D016296:Mutagenesis; D009928:Organ Specificity
keywords: CRISPR/Cas9; Drosophila; Gal4; conditional mutagenesis; gRNA