4.8 Article

Pooled genome-wide CRISPR activation screening for rapamycin resistance genes in Drosophila cells

期刊

ELIFE
卷 12, 期 -, 页码 -

出版社

eLIFE SCIENCES PUBL LTD
DOI: 10.7554/eLife.85542

关键词

CRISPR activation; genetic screening; rapamycin resistance gene; D; melanogaster

类别

向作者/读者索取更多资源

This study establishes a CRISPR activation screening platform in Drosophila cells and identifies three novel rapamycin resistance genes. Mechanistic studies reveal that one of the genes enhances resistance by activating the RTK-Akt-mTOR signaling pathway, which requires cholesterol and clathrin-coated pits at the cell membrane. This research provides a novel platform for functional genetic studies in Drosophila cells.
Loss-of-function and gain-of-function genetic perturbations provide valuable insights into gene function. In Drosophila cells, while genome-wide loss-of-function screens have been extensively used to reveal mechanisms of a variety of biological processes, approaches for performing genome-wide gain-of-function screens are still lacking. Here, we describe a pooled CRISPR activation (CRISPRa) screening platform in Drosophila cells and apply this method to both focused and genome-wide screens to identify rapamycin resistance genes. The screens identified three genes as novel rapamycin resistance genes: a member of the SLC16 family of monocarboxylate transporters (CG8468), a member of the lipocalin protein family (CG5399), and a zinc finger C2H2 transcription factor (CG9932). Mechanistically, we demonstrate that CG5399 overexpression activates the RTK-Akt-mTOR signaling pathway and that activation of insulin receptor (InR) by CG5399 requires cholesterol and clathrin-coated pits at the cell membrane. This study establishes a novel platform for functional genetic studies in Drosophila cells.

作者

我是这篇论文的作者
点击您的名字以认领此论文并将其添加到您的个人资料中。

评论

主要评分

4.8
评分不足

次要评分

新颖性
-
重要性
-
科学严谨性
-
评价这篇论文

推荐

暂无数据
暂无数据