RNA Interference (RNAi) Screening in Drosophila

Genetics: A Periodical Record of Investigations Bearing on Heredity and Variation

article

Florian Heigwer¹  Fillip Port¹  Michael Boutros¹ 

[1] Division of Signaling and Functional Genomics, German Cancer Research Center, and Department of Cell and Molecular Biology, Heidelberg University, Medical Faculty Mannheim, D-69120, Germany

关键词: Drosophila;    RNAi;    functional genomics;    high-throughput screening;    image-based screening;    bioinformatics;    genome engineering;    FlyBook;   

DOI  :  10.1534/genetics.117.300077

学科分类：医学（综合）

来源: Genetics Society of America

【 摘 要 】

In the last decade, RNA interference (RNAi), a cellular mechanism that uses RNA-guided degradation of messenger RNA transcripts, has had an important impact on identifying and characterizing gene function. First discovered in Caenorhabditis elegans , RNAi can be used to silence the expression of genes through introduction of exogenous double-stranded RNA into cells. In Drosophila , RNAi has been applied in cultured cells or in vivo to perturb the function of single genes or to systematically probe gene function on a genome-wide scale. In this review, we will describe the use of RNAi to study gene function in Drosophila with a particular focus on high-throughput screening methods applied in cultured cells. We will discuss available reagent libraries and cell lines, methodological approaches for cell-based assays, and computational methods for the analysis of high-throughput screens. Furthermore, we will review the generation and use of genome-scale RNAi libraries for tissue-specific knockdown analysis in vivo and discuss the differences and similarities with the use of genome-engineering methods such as CRISPR/Cas9 for functional analysis.

【 授权许可】

CC BY