An RNA Interference Screen for Genes Required to Shape the Anteroposterior Compartment Boundary in Drosophila Identifies the Eph Receptor
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{"title"=>"An RNA interference screen for genes required to shape the anteroposterior compartment boundary in Drosophila identifies the Eph receptor", "type"=>"journal", "authors"=>[{"first_name"=>"Daiki", "last_name"=>"Umetsu", "scopus_author_id"=>"10341169200"}, {"first_name"=>"Sebastian", "last_name"=>"Dunst", "scopus_author_id"=>"57193793358"}, {"first_name"=>"Christian", "last_name"=>"Dahmann", "scopus_author_id"=>"6602262438"}], "year"=>2014, "source"=>"PLoS ONE", "identifiers"=>{"issn"=>"19326203", "isbn"=>"1932-6203 (Electronic)\\r1932-6203 (Linking)", "sgr"=>"84916244795", "doi"=>"10.1371/journal.pone.0114340", "scopus"=>"2-s2.0-84916244795", "pui"=>"600684337", "pmid"=>"25473846"}, "id"=>"89b0884f-0b1f-35ab-ac2e-280c0e6f3eea", "abstract"=>"The formation of straight compartment boundaries separating groups of cells with distinct fates and functions is an evolutionarily conserved strategy during animal development. The physical mechanisms that shape compartment boundaries have recently been further elucidated, however, the molecular mechanisms that underlie compartment boundary formation and maintenance remain poorly understood. Here, we report on the outcome of an RNA interference screen aimed at identifying novel genes involved in maintaining the straight shape of the anteroposterior compartment boundary in Drosophila wing imaginal discs. Out of screening 3114 transgenic RNA interference lines targeting a total of 2863 genes, we identified a single novel candidate that interfered with the formation of a straight anteroposterior compartment boundary. Interestingly, the targeted gene encodes for the Eph receptor tyrosine kinase, an evolutionarily conserved family of signal transducers that has previously been shown to be important for maintaining straight compartment boundaries in vertebrate embryos. Our results identify a hitherto unknown role of the Eph receptor tyrosine kinase in Drosophila and suggest that Eph receptors have important functions in shaping compartment boundaries in both vertebrate and insect development.", "link"=>"http://www.mendeley.com/research/rna-interference-screen-genes-required-shape-anteroposterior-compartment-boundary-drosophila-identif", "reader_count"=>21, "reader_count_by_academic_status"=>{"Researcher"=>2, "Student > Doctoral Student"=>2, "Student > Ph. D. Student"=>9, "Student > Postgraduate"=>1, "Student > Master"=>1, "Student > Bachelor"=>5, "Professor"=>1}, "reader_count_by_user_role"=>{"Researcher"=>2, "Student > Doctoral Student"=>2, "Student > Ph. D. Student"=>9, "Student > Postgraduate"=>1, "Student > Master"=>1, "Student > Bachelor"=>5, "Professor"=>1}, "reader_count_by_subject_area"=>{"Biochemistry, Genetics and Molecular Biology"=>6, "Agricultural and Biological Sciences"=>14, "Physics and Astronomy"=>1}, "reader_count_by_subdiscipline"=>{"Physics and Astronomy"=>{"Physics and Astronomy"=>1}, "Agricultural and Biological Sciences"=>{"Agricultural and Biological Sciences"=>14}, "Biochemistry, Genetics and Molecular Biology"=>{"Biochemistry, Genetics and Molecular Biology"=>6}}, "reader_count_by_country"=>{"Germany"=>1, "Spain"=>1}, "group_count"=>0}

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Figshare

  • {"files"=>["https://ndownloader.figshare.com/files/1822510"], "description"=>"<p><b>A.</b> Number of RNAi lines screened categorized for the class of targeted gene products. <b>B.</b> Number of the analyzed RNAi lines and fractions of the phenotypic categories as a percentage.</p>", "links"=>[], "tags"=>["RNA interference lines", "compartment boundary formation", "compartment boundaries", "anteroposterior compartment boundary", "gene", "Drosophila wing imaginal discs", "shape compartment boundaries", "RNA interference screen", "Eph receptor tyrosine kinase"], "article_id"=>1260834, "categories"=>["Biological Sciences"], "users"=>["Daiki Umetsu", "Sebastian Dunst", "Christian Dahmann"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0114340.g003", "stats"=>{"downloads"=>5, "page_views"=>12, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Summary_of_RNA_interference_screen_/1260834", "title"=>"Summary of RNA interference screen.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-12-04 03:26:10"}
  • {"files"=>["https://ndownloader.figshare.com/files/1822492"], "description"=>"<p><b>A–C.</b> Wing discs expressing (A) no double-stranded RNA (control), (B) double-stranded RNA targeting <i>smo</i>, and (C) double-stranded RNA targeting <i>ci</i> in clones of cells marked by the expression of DsRed (red). Cells of the posterior compartment are labeled by expression of Venus under control of the <i>engrailed</i> gene (<i>en</i>-Venus, green). In (B) and (C), anterior clones along the AP boundary mis-segregate into the posterior territory of the wing disc (asterisks). Scale bar is 50 µm.</p>", "links"=>[], "tags"=>["RNA interference lines", "compartment boundary formation", "compartment boundaries", "anteroposterior compartment boundary", "gene", "Drosophila wing imaginal discs", "shape compartment boundaries", "RNA interference screen", "Eph receptor tyrosine kinase"], "article_id"=>1260816, "categories"=>["Biological Sciences"], "users"=>["Daiki Umetsu", "Sebastian Dunst", "Christian Dahmann"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0114340.g002", "stats"=>{"downloads"=>0, "page_views"=>11, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Validation_of_assay_/1260816", "title"=>"Validation of assay.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-12-04 03:26:10"}
  • {"files"=>["https://ndownloader.figshare.com/files/1822589"], "description"=>"<p><b>A.</b> Exon-intron structure of the <i>Eph</i> gene and regions targeted by the indicated RNAi lines. <b>B–D.</b> Wing imaginal discs displaying clones of cells expressing double-stranded RNA targeting <i>Eph</i> using the RNAi lines (B) 4771, (C) GL01189 or (D) GL00192. The clones of cells are identified by co-expression of DsRed (red); cells of the posterior compartment are labeled by expression of Venus under control of the <i>engrailed</i> gene (<i>en-</i>Venu<i>s</i>, green). Two clones expressing <i>Eph<sup>dsRNA</sup></i> located in different compartments sharing a common interface along the AP boundary locally distort the shape of the AP boundary (asterisks). In (B), <i>dcr2</i> is co-expressed to increase the efficiency of RNAi. <b>E.</b> Number of cases where two clones expressing double-stranded RNA targeting <i>Eph</i> located in different compartments sharing a common interface along the AP boundary distort the AP boundary towards the posterior side (A>P), towards the anterior side (P>A), or do not distort the AP boundary (no effect). Scale bar is 50 µm.</p>", "links"=>[], "tags"=>["RNA interference lines", "compartment boundary formation", "compartment boundaries", "anteroposterior compartment boundary", "gene", "Drosophila wing imaginal discs", "shape compartment boundaries", "RNA interference screen", "Eph receptor tyrosine kinase"], "article_id"=>1260878, "categories"=>["Biological Sciences"], "users"=>["Daiki Umetsu", "Sebastian Dunst", "Christian Dahmann"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0114340.g006", "stats"=>{"downloads"=>4, "page_views"=>24, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Validation_of_the_phenotype_of_Eph_RNAi_lines_/1260878", "title"=>"Validation of the phenotype of <i>Eph</i> RNAi lines.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-12-04 03:26:10"}
  • {"files"=>["https://ndownloader.figshare.com/files/1822468"], "description"=>"<p>A. Scheme of tester stock and cross to generate clones of cell expressing double-stranded RNA. The tester stock contains a <i>Flp</i> recombinase transgene under control of the heat-shock inducible promoter hsp70 (<i>hs</i>) on chromosome I <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0114340#pone.0114340-Struhl1\" target=\"_blank\">[36]</a> and the <i>engrailed-Venus</i> transgene on chromosome II (not shown). Chromosome III contains a transgene composed of the <i>Act5C</i> promoter followed by a FRT site, the coding sequence for the mouse transmembrane protein CD2 followed by a transcriptional terminator (Term), a second FRT site, and finally Gal4. We have recombined on the same chromosome a transgene expressing DsRed under control of UAS sequences (not shown). The tester stock is crossed to individual RNAi fly lines. The resulting progeny larvae are heat-shocked to express the Flp recombinase that in turn induces recombination between the two FRT sites on the tester stock. This recombination places Gal4 under the direct control of the <i>Act5C</i> promoter leading to Gal4 expression specifically in the clones of cells. Gal4 then binds to the UAS sequences in the RNAi line and directs expression of double-stranded RNA that leads to RNA interference. At the same time, Gal4 will induce expression of DsRed (not shown) in the same cells. B. Scheme of the assay. The central part of the wing imaginal disc is schematically drawn as an oval. A Venus fluorescent protein (green) is specifically expressed in all cells of the posterior compartment under control of the promoter sequences of the <i>engrailed</i> gene (<i>en</i>-Venus). Clones of cells expressing double stranded RNA are identified by co-expression of DsRed (red). Examples on the left depict wild-type clones located in the anterior (A) or posterior (P) compartments. Examples on the right depict a anterior clone mis-segregating into the posterior territory (top) and an posterior clone mis-segregating into the anterior territory (bottom) of the wing disc. Clones co-expressing DsRed and Venus are depicted in yellow.</p>", "links"=>[], "tags"=>["RNA interference lines", "compartment boundary formation", "compartment boundaries", "anteroposterior compartment boundary", "gene", "Drosophila wing imaginal discs", "shape compartment boundaries", "RNA interference screen", "Eph receptor tyrosine kinase"], "article_id"=>1260796, "categories"=>["Biological Sciences"], "users"=>["Daiki Umetsu", "Sebastian Dunst", "Christian Dahmann"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0114340.g001", "stats"=>{"downloads"=>1, "page_views"=>20, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_An_assay_to_identify_genes_required_to_maintain_the_straight_shape_of_the_AP_boundary_/1260796", "title"=>"An assay to identify genes required to maintain the straight shape of the AP boundary.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-12-04 03:26:10"}
  • {"files"=>["https://ndownloader.figshare.com/files/1822600", "https://ndownloader.figshare.com/files/1822601"], "description"=>"<div><p>The formation of straight compartment boundaries separating groups of cells with distinct fates and functions is an evolutionarily conserved strategy during animal development. The physical mechanisms that shape compartment boundaries have recently been further elucidated, however, the molecular mechanisms that underlie compartment boundary formation and maintenance remain poorly understood. Here, we report on the outcome of an RNA interference screen aimed at identifying novel genes involved in maintaining the straight shape of the anteroposterior compartment boundary in <i>Drosophila</i> wing imaginal discs. Out of screening 3114 transgenic RNA interference lines targeting a total of 2863 genes, we identified a single novel candidate that interfered with the formation of a straight anteroposterior compartment boundary. Interestingly, the targeted gene encodes for the Eph receptor tyrosine kinase, an evolutionarily conserved family of signal transducers that has previously been shown to be important for maintaining straight compartment boundaries in vertebrate embryos. Our results identify a hitherto unknown role of the Eph receptor tyrosine kinase in <i>Drosophila</i> and suggest that Eph receptors have important functions in shaping compartment boundaries in both vertebrate and insect development.</p></div>", "links"=>[], "tags"=>["RNA interference lines", "compartment boundary formation", "compartment boundaries", "anteroposterior compartment boundary", "gene", "Drosophila wing imaginal discs", "shape compartment boundaries", "RNA interference screen", "Eph receptor tyrosine kinase"], "article_id"=>1260887, "categories"=>["Biological Sciences"], "users"=>["Daiki Umetsu", "Sebastian Dunst", "Christian Dahmann"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0114340.s001", "https://dx.doi.org/10.1371/journal.pone.0114340.s002"], "stats"=>{"downloads"=>5, "page_views"=>14, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/An_RNA_Interference_Screen_for_Genes_Required_to_Shape_the_Anteroposterior_Compartment_Boundary_in_Drosophila_Identifies_the_Eph_Receptor/1260887", "title"=>"An RNA Interference Screen for Genes Required to Shape the Anteroposterior Compartment Boundary in <i>Drosophila</i> Identifies the Eph Receptor", "pos_in_sequence"=>0, "defined_type"=>4, "published_date"=>"2014-12-04 03:26:10"}
  • {"files"=>["https://ndownloader.figshare.com/files/1822580"], "description"=>"<p><b>A–C.</b> Wing imaginal discs displaying clones of cells expressing double-stranded RNA targeting (A–B) <i>Eph</i> or (C) <i>FasIII.</i> The clones of cells are identified by co-expression of DsRed (red); cells of the posterior compartment are labeled by expression of Venus under control of the <i>engrailed</i> gene (<i>en-</i>Venus, green). (A) Individual clones abutting the AP boundary from either the A or P compartment do not influence the shape of the boundary (asterisk). (B) Two clones expressing <i>Eph<sup>dsRNA</sup></i> (ID-Number 4771 (VDRC)) located in different compartments sharing a common interface along the AP boundary locally distort the shape of the AP boundary (asterisks). (C) Two clones expressing <i>FasIII<sup>dsRNA</sup></i> located in different compartments sharing a common interface along the AP boundary do not distort the shape of the AP boundary (asterisks). Scale bar is 50 µm.</p>", "links"=>[], "tags"=>["RNA interference lines", "compartment boundary formation", "compartment boundaries", "anteroposterior compartment boundary", "gene", "Drosophila wing imaginal discs", "shape compartment boundaries", "RNA interference screen", "Eph receptor tyrosine kinase"], "article_id"=>1260869, "categories"=>["Biological Sciences"], "users"=>["Daiki Umetsu", "Sebastian Dunst", "Christian Dahmann"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0114340.g005", "stats"=>{"downloads"=>0, "page_views"=>22, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Clones_of_cells_expressing_double_stranded_RNA_targeting_the_Eph_gene_distort_the_AP_boundary_/1260869", "title"=>"Clones of cells expressing double-stranded RNA targeting the <i>Eph</i> gene distort the AP boundary.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-12-04 03:26:10"}
  • {"files"=>["https://ndownloader.figshare.com/files/1822547"], "description"=>"<p><b>A–F.</b> Wing imaginal discs displaying clones of cells expressing double-stranded RNA targeting (A) <i>α-cat</i>, (B) <i>wts</i>, (C) <i>PlexA</i>, (D) <i>ph-p</i>, (E) <i>kto</i>, and (F) <i>mad</i>. The clones of cells are identified by co-expression of DsRed (red); cells of the posterior compartment are labeled by expression of Venus under control of the <i>engrailed</i> gene (<i>en</i>-Venus, green). Clones of cells distorting the AP boundary are marked by an asterisk. Scale bars are 50 µm.</p>", "links"=>[], "tags"=>["RNA interference lines", "compartment boundary formation", "compartment boundaries", "anteroposterior compartment boundary", "gene", "Drosophila wing imaginal discs", "shape compartment boundaries", "RNA interference screen", "Eph receptor tyrosine kinase"], "article_id"=>1260854, "categories"=>["Biological Sciences"], "users"=>["Daiki Umetsu", "Sebastian Dunst", "Christian Dahmann"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0114340.g004", "stats"=>{"downloads"=>0, "page_views"=>17, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Examples_of_phenotypes_observed_in_the_RNA_interference_screen_/1260854", "title"=>"Examples of phenotypes observed in the RNA interference screen.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-12-04 03:26:10"}

PMC Usage Stats | Further Information

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Relative Metric

{"start_date"=>"2014-01-01T00:00:00Z", "end_date"=>"2014-12-31T00:00:00Z", "subject_areas"=>[{"subject_area"=>"/Biology and life sciences", "average_usage"=>[291]}, {"subject_area"=>"/Biology and life sciences/Cell biology", "average_usage"=>[286]}, {"subject_area"=>"/Biology and life sciences/Developmental biology", "average_usage"=>[285]}, {"subject_area"=>"/Biology and life sciences/Genetics", "average_usage"=>[306, 482]}, {"subject_area"=>"/Biology and life sciences/Organisms", "average_usage"=>[310]}]}
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