RNA Interference Screening for Genes Regulating Drosophila Muscle Morphogenesis
- PMID: 30367424
- DOI: 10.1007/978-1-4939-8897-6_20
RNA Interference Screening for Genes Regulating Drosophila Muscle Morphogenesis
Abstract
RNA interference (RNAi) is the method of choice to systematically test for gene function in an intact organism. The model organism Drosophila has the advantage that RNAi is cell autonomous, meaning it does not spread from one cell to the next. Hence, RNAi can be performed in a tissue-specific manner by expressing short or long inverted repeat constructs (hairpins) designed to target mRNAs from one specific target gene. This achieves tissue-specific knock-down of a target gene of choice. Here, we detail the methodology to test gene function in Drosophila muscle tissue by expressing hairpins in a muscle-specific manner using the GAL4-UAS system. We further discuss the systematic RNAi resource collections available which also permit large scale screens in a muscle-specific manner. The full power of such screens is revealed by combination of high-throughput assays followed by detailed morphological assays. Together, this chapter should be a practical guide to enable the reader to either test a few candidate genes, or large gene sets for particular functions in Drosophila muscle tissue and provide first insights into the biological process the gene might be important for in muscle.
Keywords: Drosophila; GAL4-UAS; Genetic screens; Muscle; Off-target effects; RNAi; Sarcomere.
Similar articles
-
RNA Interference (RNAi) Screening in Drosophila.Genetics. 2018 Mar;208(3):853-874. doi: 10.1534/genetics.117.300077. Genetics. 2018. PMID: 29487145 Free PMC article.
-
A Guide to Genome-Wide In Vivo RNAi Applications in Drosophila.Methods Mol Biol. 2016;1478:117-143. doi: 10.1007/978-1-4939-6371-3_6. Methods Mol Biol. 2016. PMID: 27730578 Review.
-
Design and Methods of Large-Scale RNA Interference Screens in Drosophila.Methods Mol Biol. 2016;1470:163-9. doi: 10.1007/978-1-4939-6337-9_13. Methods Mol Biol. 2016. PMID: 27581292
-
The purine synthesis gene Prat2 is required for Drosophila metamorphosis, as revealed by inverted-repeat-mediated RNA interference.Genetics. 2006 Mar;172(3):1621-31. doi: 10.1534/genetics.105.045641. Epub 2005 Dec 1. Genetics. 2006. PMID: 16322507 Free PMC article.
-
A guide to study Drosophila muscle biology.Methods. 2014 Jun 15;68(1):2-14. doi: 10.1016/j.ymeth.2014.02.037. Epub 2014 Mar 12. Methods. 2014. PMID: 24625467 Review.
Cited by
-
A Candidate RNAi Screen Reveals Diverse RNA-Binding Protein Phenotypes in Drosophila Flight Muscle.Cells. 2021 Sep 22;10(10):2505. doi: 10.3390/cells10102505. Cells. 2021. PMID: 34685485 Free PMC article.
-
Muscle degeneration in aging Drosophila flies: the role of mechanical stress.Skelet Muscle. 2024 Aug 20;14(1):20. doi: 10.1186/s13395-024-00352-4. Skelet Muscle. 2024. PMID: 39164781 Free PMC article.
-
Reduction of Drosophila Mitochondrial RNase P in Skeletal and Heart Muscle Causes Muscle Degeneration, Cardiomyopathy, and Heart Arrhythmia.Front Cell Dev Biol. 2022 May 19;10:788516. doi: 10.3389/fcell.2022.788516. eCollection 2022. Front Cell Dev Biol. 2022. PMID: 35663400 Free PMC article.
-
Cofilin Loss in Drosophila Muscles Contributes to Muscle Weakness through Defective Sarcomerogenesis during Muscle Growth.Cell Rep. 2020 Jul 21;32(3):107893. doi: 10.1016/j.celrep.2020.107893. Cell Rep. 2020. PMID: 32697999 Free PMC article.
-
Rbfox1 is required for myofibril development and maintaining fiber type-specific isoform expression in Drosophila muscles.Life Sci Alliance. 2022 Jan 7;5(4):e202101342. doi: 10.26508/lsa.202101342. Print 2022 Apr. Life Sci Alliance. 2022. PMID: 34996845 Free PMC article.
MeSH terms
Substances
LinkOut - more resources
Full Text Sources
Molecular Biology Databases
Research Materials