Identification of Split-GAL4 Drivers and Enhancers That Allow Regional Cell Type Manipulations of the Drosophila melanogaster Intestine
Identification of Split-GAL4 Drivers and Enhancers That Allow Regional Cell Type Manipulations of the Drosophila melanogaster Intestine
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DOI：
10.1534/genetics.120.303625
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发表时间：
2020-12-01
期刊：
影响因子：
3.3
通讯作者：
Sokol, Nicholas S.
中科院分区：
文献类型：
--
作者：
Ariyapala, Ishara S.;Holsopple, Jessica M.;Sokol, Nicholas S.
果蝇成虫中肠是由几种具有不同区域特性的主要细胞类型组成的模式上皮组织。其分析的局限性之一是缺乏基于这些区域身份操纵基因表达的工具。为了克服这一障碍，我们将交叉分裂GAL 4系统应用于成年中肠，并报告了653种按区域和细胞类型标记细胞的驱动程序组合。我们首先从筛选的7300个驱动程序中鉴定出424个具有中肠表达的分裂-GAL 4驱动程序，然后在与三个参考驱动程序配对时评估这424个驱动程序中每一个的表达模式，所述参考驱动程序特异性地报告祖细胞、肠内分泌细胞或肠细胞中的活性。我们还使用另一种参考驱动因子评估了在祖细胞中表达的驱动因子的子集在成肠细胞中的表达。我们表明，驱动程序组合可以定义新的细胞群体，通过识别一个驱动程序，标志着一个独特的子集的肠内分泌细胞表达基因通常与祖细胞。与整套驱动程序组合相关的区域细胞类型模式记录在一个免费提供的网站上，为设计数千种额外的驱动程序组合提供信息，以实验性地操纵小肠细胞的子集。此外，我们表明，肠道增强子确定与分裂GAL 4系统可以赋予其他转基因记者等效的表达模式。总之，这里报告的资源将使更精确的靶向基因表达研究肠过程，上皮细胞功能和影响自我更新组织的疾病。
The Drosophila adult midgut is a model epithelial tissue composed of a few major cell types with distinct regional identities. One of the limitations to its analysis is the lack of tools to manipulate gene expression based on these regional identities. To overcome this obstacle, we applied the intersectional split-GAL4 system to the adult midgut and report 653 driver combinations that label cells by region and cell type. We first identified 424 split-GAL4 drivers with midgut expression from similar to 7300 drivers screened, and then evaluated the expression patterns of each of these 424 when paired with three reference drivers that report activity specifically in progenitor cells, enteroendocrine cells, or enterocytes. We also evaluated a subset of the drivers expressed in progenitor cells for expression in enteroblasts using another reference driver. We show that driver combinations can define novel cell populations by identifying a driver that marks a distinct subset of enteroendocrine cells expressing genes usually associated with progenitor cells. The regional cell type patterns associated with the entire set of driver combinations are documented in a freely available website, providing information for the design of thousands of additional driver combinations to experimentally manipulate small subsets of intestinal cells. In addition, we show that intestinal enhancers identified with the split-GAL4 system can confer equivalent expression patterns on other transgenic reporters. Altogether, the resource reported here will enable more precisely targeted gene expression for studying intestinal processes, epithelial cell functions, and diseases affecting self-renewing tissues.