Open AccessSilencing of genes in cultured Drosophila neurons by RNA interference
Author(s) -
Shail K. Sharma,
Marshall W. Nirenberg
Publication year - 2007
Publication title -
proceedings of the national academy of sciences
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 5.011
H-Index - 771
eISSN - 1091-6490
pISSN - 0027-8424
DOI - 10.1073/pnas.0704299104
Subject(s) - rna interference , gene silencing , biology , rna silencing , transfection , microbiology and biotechnology , small interfering rna , neuroblast , schneider 2 cells , neurite , gene , gene knockdown , synapsin i , nucleofection , rna , genetics , neurogenesis , synaptic vesicle , vesicle , membrane , in vitro
Cultures of neuroblasts that generate abundant neurons were established fromDrosophila embryos to study silencing of genes by RNA interference (RNAi). Cultured cells expressed ELAV, a marker of neurons, Futsch, a marker of neurites, and Synapsin, Synaptobrevin, and Synaptogamin, proteins involved in neurotransmitter secretion. Conditions were found for efficient transfection of cells with siRNAs for ELAV or the insulin-like receptor, which resulted in marked decreases in neurons that express ELAV and Futsch. Cells also were successfully transfected with long-chain Sox-Neuro dsRNA resulting in a 55% reduction of neurons expressing Futsch. The results suggest that this cultured neural cell system can be used to study RNAi-dependent silencing of genes involved in many kinds of neural functions.
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