Key resources table.
| Reagent type (species) or resource |
Designation | Source or reference |
Identifiers | Additional information |
|---|---|---|---|---|
| Strain, strain background (Escherichia coli) | 5-alpha competent E. coli | NEB | C2987I | High Efficiency chemically competent |
| Genetic reagent (Drosophila melanogaster) | cnnf04547 | Exelixis at Harvard Medical School | Exelixis:f04547; Flybase:FBst1019233 | FlyBase symbol: cnnf04547 ; Stock discontinued at the Exelixis and available from Conduit lab upon request. |
| Genetic reagent (D. melanogaster) | plp5 | Bloomington Drosophila Stock Center | BDSC:9567; FLYB:FBst0009567; RRID:BDSC_9567 | FlyBase symbol: ru1 Plp5 st1/TM6C, Sb1 Tb1 |
| Genetic reagent (D. melanogaster) | plps2172 | Bloomington Drosophila Stock Center | BDSC:12089; Flybase:FBst0012089; RRID:BDSC_12089 | FlyBase symbol: ‘w[1118]; P{w[+mC]=lacW}Plp[s2172]/TM3, Sb[1]’ |
| Genetic reagent (D. melanogaster) | UAS-mCD8-RFP | Bloomington Drosophila Stock Center | BDSC:27392; Flybase:FBti0115769; RRID:BDSC_27392 | FlyBase symbol: ‘w[*]; P{w[+mC]=UAS-mCD8.ChRFP}3’ |
| Genetic reagent (D. melanogaster) | UAS-EB1-GFP | Bloomington Drosophila Stock Center | BDSC:35512; Flybase:FBti0141213; RRID:BDSC_35512 | FlyBase symbol: ‘w[*]; P{w[+mC]=UAS-EB1-GFP}3’ |
| Genetic reagent (D. melanogaster) | UAS-ManII-GFP | Bloomington Drosophila Stock Center | BDSC:65248; Flybase:FBti018367; RRID:BDSC_65248 | FlyBase symbol: ‘w[1118]; P{w[+mC]=UAS-ManII-EGFP}2; TM2/TM6B, Tb[1]’ |
| Genetic reagent (D. melanogaster) | UAS-ManII-mCherry | Ori-McKenney et al., 2012 (doi:10.1016/j.neuron.2012.10.008) | Fly genotype: ‘w[1118]; P{w[+mC]=UAS-ManII-mCherry}/CyO’; gift from Yuh-Nung Jan | |
| Genetic reagent (D. melanogaster) | ppk-Gal4 | Bloomington Drosophila Stock Center | BDSC:32078; Flybase:FBti0127690; RRID:BDSC_32078 | FlyBase symbol: ‘w[*]; P{w[+mC]=ppk-GAL4.G}2’ |
| Genetic reagent (D. melanogaster) | ppk-Gal4 | Bloomington Drosophila Stock Center | BDSC:32079; Flybase:FBti013120; RRID:BDSC_32079 | FlyBase symbol: ‘w[*]; P{w[+mC]=ppk-GAL4.G}3’ |
| Genetic reagent (D. melanogaster) | 221-Gal4 | Bloomington Drosophila Stock Center | BDSC:26259; Flybase:FBti011433; RRID:BDSC_26259 | FlyBase symbol: ‘w[*]; Pin[1]/CyO; P{GawB}221w-’ |
| Genetic reagent (D. melanogaster) | ppkCD4-tdGFP | Bloomington Drosophila Stock Center | BDSC:35842; Flybase:FBti014342; RRID:BDSC_35842 | FlyBase symbol: ‘w[1118]; P{w[+mC]=ppk-CD4-tdGFP}1b’ |
| Genetic reagent (D. melanogaster) | UAS-Dicer 2 | Bloomington Drosophila Stock Center | BDSC:24646; Flybase:FBti010143; RRID:BDSC_24646 | FlyBase symbol: P{w[+mC]=UAS-Dcr2.D}1,w[1118] |
| Genetic reagent (D. melanogaster) | γ-tubulin37C RNAi; UAS-γ-tubulin37C RNAi | Bloomington Drosophila Stock Center | BDSC:32513; Flybase:FBti0132207; RRID:BDSC_32513 | FlyBase symbol: ‘y[1] sc[*] v[1] sev[21]; P{y[+t7.7] v[+t1.8]=TRiP.HMS00517}attP2’ |
| Genetic reagent (D. melanogaster) | Kap3 RNAi; UAS-Kap3 RNAi | ViennaDrosophila Resource Center | FBst0466097; VDRC ID:45400 | FlyBase symbol: ‘w1118;P{GD7377}v45400’ |
| Genetic reagent (D. melanogaster) | Klp64D RNAi | ViennaDrosophila Resource Center | FBst0466080; VDRC ID:45373 | FlyBase symbol: ‘w1118;P{GD7048}v45373’ |
| Genetic reagent (D. melanogaster) | γ-tubulin23c-sfGFP | Tovey and Conduit, 2018 (https://doi.org/10.1016/j.cub.2018.05.044) | insertion of sfGFP with a 4X GlyGlySer linker at the C-terminus | |
| Genetic reagent (D. melanogaster) | γ-tubulin23c-eGFP | This paper | insertion of eGFP with a 4X Ser linker at the C-terminus. Generated by inDroso | |
| Genetic reagent (D. melanogaster) | sfGFP-Cnn-P1 | This paper | insertion of sfGFP with a 4X GlyGlySer linker at the start of exon 1a | |
| Genetic reagent (D. melanogaster) | sfGFP-Cnn-P2 | This paper | insertion of sfGFP with a 4X GlyGlySer linker at the start of exon 1b | |
| Antibody | anti-GFP (mouse monoclonal) | Roche | Cat# 11814460001; RRID:AB_390913 | IF(1:250) |
| Antibody | anti-Cnn (rabbit polyclonal) | Lucas and Raff, 2007 (https://doi.org/10.1083/jcb.200704081) | IF(1:1000); gift from Jordan Raff | |
| Antibody | anti-Plp (rabbit polyclonal) | Martinez-Campos et al., 2004 (https://doi.org/10.1083/jcb.200402130) | IF(1:500); gift from Jordan Raff | |
| Antibody | anti-Arl1 (chicken polyclonal) | Torres et al., 2014 (doi:10.1242/jcs.122028) | IF(1:200); gift from Sean Munroe | |
| Antibody | anti-GM130 (rabbit polyclonal) | Abcam | Cat# ab30637; RRID:AB_732675 | IF(1:300) |
| Antibody | Alexa-647 conjugated Hrp (goat polyclonal) | Jackson | Jackson ImmunoResearch 123-605-021; RRID:AB_2338967 | IF(1:500) |
| Antibody | ATTO-488 GFP-booster (Camelid single domain ‘nanobody’) |
Chromotek | gb2AF488-10; RRID:AB_2827573 | IF(1:200) |
| Antibody | anti-Mouse IgG H and L Alexa Fluor 488 (goat polyclonal) | Abcam | Cat# ab150117, RRID:AB_2688012 | IF(1:500) |
| Antibody | anti-Rabbit IgG H and L Alexa Fluor 568 (goat polyclonal) | Thermo Fisher Scientific | Cat# A-11036, RRID:AB_10563566 | IF(1:500) |
| Recombinant DNA reagent | pCDF3 (plasmid) | Addgene | RRID:Addgene_49410 | pCFD3-dU6:3gRNA; gift from Simon Bullock |
| Recombinant DNA reagent | pBluescript SK+ (plasmid) | Simon Bullock lab | Standard cloning vector; gift from Simon Bullock | |
| Recombinant DNA reagent | pBS-KS-attB1-2-PT-SA-SD-2-sfGFP-FIAsh-StrepII-TEV-3x-FLAG (plasmid) | Drosophila Genomics Resource Center | Stock number:1314 | Vector used to amplify sf GFP sequence |
| Sequence-based reagent | CnnNt_P1T2_f | This paper | Guide oligo | GTCGTGTTTAGACTGGTCCATGGG To generate Bbs1 cleavable fragment to clone into Bbs1 cut pCFD3 for Cnn promoter one tagging. Guide sequence underlined. |
| Sequence-based reagent | CnnNt_P1T2_b | This paper | Guide oligo | AAACCCCATGGACCAGTCTAAACA; To generate Bbs1 cleavable fragment to clone into Bbs1 cut pCFD3 for Cnn promoter one tagging. Guide sequence underlined. |
| Sequence-based reagent | CnnNt_P2T2_f | This paper | Guide oligo | GTCGTTAAATGAAACATAGAATA; To generate Bbs1 cleavable fragment to clone into Bbs1 cut pCFD3 for Cnn promoter two tagging. Guide sequence underlined. |
| Sequence-based reagent | CnnNt_P2T2_b | This paper | Guide oligo | AAACTATTCTATGTTTCATTTAA; To generate Bbs1 cleavable fragment to clone into Bbs1 cut pCFD3 for Cnn promoter two tagging. Guide sequence underlined. |
| Sequence-based reagent | cnn_UHA_P1_GFP_f | This paper | PCR primers | AAAGTTAACTATTTGAGGACCTCCCATGGTGTCCAAGGGCGAGGAG; used to amplify sfGFP with overhangs for HiFi cloning into pBS containing homology arms for for Cnn promoter 1 |
| Sequence-based reagent | cnn_DHA_P1_GFP_b | This paper | PCR primers | CCCGCAAAACCTGTTTAGACTGGTCGGATCCGCCGCTACCTCCGCTTCCACCGGAACCTCCCTTGTACAGCTCATCCATGCC; used to amplify sfGFP with overhangs for HiFi cloning into pBS containing homology arms for for Cnn promoter 1. Also contains sequence for 4X GlyGlySer linker |
| Sequence-based reagent | cnn_UHA_P2_GFP_f | This paper | PCR primers | GCAAATGTTAAATGAAAGATACAATATGGTGTCCAAGGGCGAGGAG; used to amplify sfGFP with overhangs for HiFi cloning into pBS containing homology arms for for Cnn promoter 2 |
| Sequence-based reagent | cnn_DHA_P2_GFP_b | This paper | PCR primers | GTGAGGTAGATCGAAAGATACCCGCGGATCCGCCGCTACCTCCGCTTCCACCGGAACCTCCCTTGTACAGCTCATCCATGCC; used to amplify sfGFP with overhangs for HiFi cloning into pBS containing homology arms for for Cnn promoter 2. Also contains sequence for 4X GlyGlySer linker |
| Sequence-based reagent | cnn_UHA_P1_f | This paper | PCR primers | GAAAGCTAAGCAGATTCTTCAGC; used to amplify upstream homology arm for promoter 1 |
| Sequence-based reagent | cnn_UHA_P1_b | This paper | PCR primers | GGGAGGTCCTCAAATAGTTAAC; used to amplify upstream homology arm for promoter 1 |
| Sequence-based reagent | cnn_DHA_P1_f | This paper | PCR primers | GACCAGTCTAAACAGGTTTTGC; used to amplify downstream homology arm for promoter 1 |
| Sequence-based reagent | cnn_DHA_P1_b | This paper | PCR primers | GCTCTCTGCACGTCCAAATAAAC; used to amplify downstream homology arm for promoter 1 |
| Sequence-based reagent | cnn_UHA_P2_f | This paper | PCR primers | CACTCGCTATAGTCTCGATCC; used to amplify upstream homology arm for promoter 2 |
| Sequence-based reagent | cnn_UHA_P2_b | This paper | PCR primers | ATTGTATCTTTCATTTAACATTTGCGCTC; used to amplify upstream homology arm for promoter 2 |
| Sequence-based reagent | cnn_DHA_P2_f | This paper | PCR primers | GCGGGTATCTTTCGATCTACC; used to amplify downstream homology arm for promoter 2 |
| Sequence-based reagent | cnn_DHA_P2_b | This paper | PCR primers | TCCCGTGGAACAGGAACCAC; used to amplify downstream homology arm for promoter 2 |
| Sequence-based reagent | cnn_DHA_P1_pBS_f | This paper | PCR primers | CGGTTTATTTGGACGTGCAGAGAGCGGGGGATCCACTAGTTCTAGAG; used to amplify pBS backbone for promoter 1. Contains overhangs for HiFi cloning. |
| Sequence-based reagent | cnn_UHA_P1_pBS_b | This paper | PCR primers | AAGCTGAAGAATCTGCTTAGCTTTCGGGCTGCAGGAATTCGATATC; used to amplify pBS backbone for promoter 1. Contains overhangs for HiFi cloning. |
| Sequence-based reagent | cnn_DHA_P2_pBS_f | This paper | PCR primers | TTGCTGTGGTTCCTGTTCCACGGGAGGGGGATCCACTAGTTCTAGAG; used to amplify pBS backbone for promoter 2. Contains overhangs for HiFi cloning. |
| Sequence-based reagent | cnn_UHA_P2_pBS_b | This paper | PCR primers | AAGTGGATCGAGACTATAGCGAGTGGGGCTGCAGGAATTCGATATC; used to amplify pBS backbone for promoter 2. Contains overhangs for HiFi cloning. |
| Other | Live imaging solution | Thermo Fisher Scientific | A14291DJ | Larvae imaging |