Identification of neural outgrowth genes using genome-wide RNAi.

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dc.contributor.author Sepp, Katharine J
dc.contributor.author Hong, Pengyu
dc.contributor.author Lizarraga, Sofia B
dc.contributor.author Liu, Judy S
dc.contributor.author Mejia, Luis A
dc.contributor.author Walsh, Christopher A
dc.contributor.author Perrimon, Norbert
dc.date.accessioned 2019-02-05T18:24:27Z
dc.date.available 2019-02-05T18:24:27Z
dc.date.issued 2008
dc.identifier.issn 1553-7404
dc.identifier.other PMC2435276
dc.identifier.uri https://hdl.handle.net/10192/36497
dc.description.abstract While genetic screens have identified many genes essential for neurite outgrowth, they have been limited in their ability to identify neural genes that also have earlier critical roles in the gastrula, or neural genes for which maternally contributed RNA compensates for gene mutations in the zygote. To address this, we developed methods to screen the Drosophila genome using RNA-interference (RNAi) on primary neural cells and present the results of the first full-genome RNAi screen in neurons. We used live-cell imaging and quantitative image analysis to characterize the morphological phenotypes of fluorescently labelled primary neurons and glia in response to RNAi-mediated gene knockdown. From the full genome screen, we focused our analysis on 104 evolutionarily conserved genes that when downregulated by RNAi, have morphological defects such as reduced axon extension, excessive branching, loss of fasciculation, and blebbing. To assist in the phenotypic analysis of the large data sets, we generated image analysis algorithms that could assess the statistical significance of the mutant phenotypes. The algorithms were essential for the analysis of the thousands of images generated by the screening process and will become a valuable tool for future genome-wide screens in primary neurons. Our analysis revealed unexpected, essential roles in neurite outgrowth for genes representing a wide range of functional categories including signalling molecules, enzymes, channels, receptors, and cytoskeletal proteins. We also found that genes known to be involved in protein and vesicle trafficking showed similar RNAi phenotypes. We confirmed phenotypes of the protein trafficking genes Sec61alpha and Ran GTPase using Drosophila embryo and mouse embryonic cerebral cortical neurons, respectively. Collectively, our results showed that RNAi phenotypes in primary neural culture can parallel in vivo phenotypes, and the screening technique can be used to identify many new genes that have important functions in the nervous system.
dc.format.extent 1 file
dc.language English
dc.language.iso eng
dc.relation.isversionof https://dx.doi.org/10.1371/journal.pgen.1000111
dc.rights Creative Commons Attribution 4.0 International License
dc.rights.uri http://creativecommons.org/licenses/by/4.0/
dc.subject Animals
dc.subject Cells, Cultured
dc.subject Drosophila
dc.subject Genome
dc.subject Genomics
dc.subject Mice
dc.subject Nervous System
dc.subject Neurons
dc.subject Phenotype
dc.subject RNA Interference
dc.subject RNA, Small Interfering
dc.subject ran GTP-Binding Protein
dc.subject RNA, Small Interfering
dc.subject Ran protein, mouse
dc.subject ran GTP-Binding Protein
dc.title Identification of neural outgrowth genes using genome-wide RNAi.
dc.type Article
dc.contributor.department Department of Computer Science
dc.relation.journal PLoS Genetics
dc.identifier.pmid 18604272


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