Silencing of genes in cultured Drosophila neurons by RNA interference.
Cultures of neuroblasts that generate abundant neurons were established from Drosophila 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.
Sharma
Shail K
SK
Laboratory of Biochemical Genetics, National Heart, Lung, and Blood Institute, National Institutes of Health, Bethesda, MD 20892, USA.
Nirenberg
Marshall
M
D000818:Animals; D015415:Biomarkers; D002454:Cell Differentiation; D048430:Cell Shape; D002478:Cells, Cultured; D029721:Drosophila Proteins / Q000235:genetics / Q000378:metabolism; D004331:Drosophila melanogaster / Q000166:cytology / Q000235:genetics* / Q000378:metabolism; D051959:ELAV Proteins / Q000235:genetics / Q000378:metabolism; D018507:Gene Expression Regulation, Developmental; D017344:Genes, Insect / Q000235:genetics*; D008869:Microtubule-Associated Proteins / Q000378:metabolism; D009414:Nerve Growth Factors / Q000378:metabolism; D009474:Neurons / Q000166:cytology / Q000378:metabolism*; D034622:RNA Interference; D011972:Receptor, Insulin / Q000235:genetics; D013997:Time Factors
D015415:Biomarkers; D029721:Drosophila Proteins; D051959:ELAV Proteins; C498009:ELAV protein, Drosophila; D008869:Microtubule-Associated Proteins; D009414:Nerve Growth Factors; C409635:futsch protein, Drosophila; D011972:Receptor, Insulin