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The His2Av-mRFP is a fluorescent protein that can be used to label histone proteins in Drosophila cells. It consists of the Drosophila histone variant His2Av fused to the monomeric red fluorescent protein (mRFP). This fusion protein allows for the visualization of chromatin dynamics and nuclear organization in living Drosophila cells.

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Lab products found in correlation

Mhc gal4, by Bloomington Drosophila Stock Center (1 mentions) Np6293 gal4, by Kyoto Stock Center (1 mentions) Uas sima rnai, by Bloomington Drosophila Stock Center (1 mentions) Nrv2 gal4, by Bloomington Drosophila Stock Center (1 mentions) Elav gal4, by Bloomington Drosophila Stock Center (1 mentions) Repo gal4, by Bloomington Drosophila Stock Center (1 mentions) Nos gal4, by Bloomington Drosophila Stock Center (1 mentions) His2av egfp, by Bloomington Drosophila Stock Center (1 mentions) Sqh gfp, by Bloomington Drosophila Stock Center (1 mentions)

5 protocols using his2av mrfp

1

Drosophila Genetics and Cell Imaging

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All D. melanogaster stocks were cultured on standard cornmeal-yeast medium. The following fly strains were used in this study: w1118 Canton-S (wild type), His2Av-mRFP (Bloomington stock number 23650 or 23651), mRFP-HP1a (30562), eyeless-gal4 (5535), maternal tubulin-gal4 (7063), UAS-shRNAcdc7 (TRiP GL00585), mei41D3 (gift from Tin Tin Su, University of Colorado Boulder), mCherry-PCNA (this study), rif1-EGFP (this study), cdc7-EGFP (this study), rif1- (this study), UASp>Rif1-3xFlag (this study), UASp>Rif1-15a-3xFlag (this study).
Seller C.A, & O’Farrell P.H. (2018). Rif1 prolongs the embryonic S phase at the Drosophila mid-blastula transition. PLoS Biology, 16(5), e2005687.
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2

Fruit Fly Stocks and Transgenics

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Stocks of Drosophila melanogaster were cultured on standard cornmeal–yeast medium. The following previously described stocks were used in this study: w1118 Canton-S (wild type), His2Av-mRFP (Bloomington stock no. 23650), EGFP-HP1a (30561), maternal tubulin-Gal4 (7063), sfGFP-zelda (M. Harrison), ORC2-GFP; orc21/y4 (M. Gossen), G9aRG5 (P. Spierer), grp06034, Su(var)3–906, Su(var)3–917, and RIF1-EGFP.
Embryo injections for the transgenic lines described in this study were done by Rainbow Transgenic Flies. ΦC31-mediated integration generated the JabbaTrap transgene UASp-VhhGFP4-jabba-VhhGFP4attP-9A. The following stocks were created using CRISPR–Cas9 gene editing: sfGFP-Su(z)12, sfGFP-G9a, EGFP-egg, and Halo-egg. See Supplemental Figure S1 for details of endogenous tagging. Following generation, modified flies were crossed to our laboratory's wild-type stock (w1118 Canton-S) for at least five generations. Plasmid DNA and sequence files are available on request.
Seller C.A., Cho C.Y, & O'Farrell P.H. (2019). Rapid embryonic cell cycles defer the establishment of heterochromatin by Eggless/SetDB1 in Drosophila. Genes & Development, 33(7-8), 403-417.
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3

Visualizing Transcription Dynamics in Drosophila

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Embryos were obtained from crosses between males carrying MS2 reporters and females carrying the maternally expressed MCP-NoNLS-eGFP (Garcia et al., 2013 (link)) and His2Av-mRFP (Bloomington # 23561) transgenes both on the second chromosome. Embryos with reduced activity of Bcd were obtained from females which were in addition heterozygotes for the bcdΔ molecular null allele or for the bcdE1 allele (bcd6). Unless otherwise specified, all MS2 reporters were inserted at the vk33 docking site (Bloomington # 9750) via φC31-mediated integration system (Venken et al., 2006 (link)) by BestGene. The site of insertion was chosen because the transcription dynamics of the original hb-P2 reporter (Lucas et al., 2018 (link)) inserted at this site was indistinguishable from the transcription dynamics of two randomly inserted siblings (Figure 1—figure supplement 1, panels A-C). All fly stocks were maintained at 25°C.
Fernandes G., Tran H., Andrieu M., Diaw Y., Perez Romero C., Fradin C., Coppey M., Walczak A.M, & Dostatni N. (2022). Synthetic reconstruction of the hunchback promoter specifies the role of Bicoid, Zelda and Hunchback in the dynamics of its transcription. eLife, 11, e74509.
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4

Drosophila Genetic Toolkit Utilization

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All flies were reared at 25°C. w1118 was used as wild-type control and to backcross with trh1 or trh2. The sources of fly strains are as follows: trh2, elav-GAL4, MHC-GAL4, repo-GAL4, UAS-trh, UAS-sima, UAS-sima-RNAi, Nrv2-GAL4, and His2Av-mRFP were obtained from Bloomington Drosophila Stock Center (BDSC); trh1, NP6293-GAL4, and UAS-wg-RNAi from Kyoto Stock Center; and UAS-trh-RNAi, UAS-fga-RNAi, and UAS-fz2-RNAi from Vienna Drosophila Resource Center (VDRC). Also used were btl-GAL4[56 ], moody-GAL4 [57 (link)], alrm-GAL4[58 (link)], UAS-wg, and GFP-ODD [29 (link)]. The repo-cyto-GFP line was generated with the sequence for cytoplasmic GFP under the control of the 4.3 kb repo promoter, which recapitulates the full repo expression pattern.
Chen P.Y., Tsai Y.W., Cheng Y.J., Giangrande A, & Chien C.T. (2019). Glial response to hypoxia in mutants of NPAS1/3 homolog Trachealess through Wg signaling to modulate synaptic bouton organization. PLoS Genetics, 15(8), e1007980.
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5

Drosophila Imaging Toolkit Protocols

Cited 1 time
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The following fly strains were obtained from the Bloomington Drosophila Stock Center: nos-Gal4 (#7303), his2av::mRFP (#23651), his2av::eGFP (#24163), sti shRNA (#35392), sep2::GFP (#26257), sfGFP::sti (#81646), sqh::GFP (#57145), mRFP::anillin (#52220), shrub::GFP (#32559) and sfGFP::tum (#76264). We imaged Pavarotti (Pav) at endogenous levels with Pav::GFP22 (link) or with Pav::mCherry (this study). uasp-GFP-alix44 (link) flies were obtained from Kaisa Haglund (Oslo University Hospital) and uas-spas-eGFP51 (link) flies from Damien Brünner (University of Zurich). 20XUAS-mC3PA-GFP was provided by Pfeiffer et al.52 (link). We examined the behavior of microtubules with a GFP-tagged microtubule-binding protein Jupiter::GFP53 (link),54 (link) using the Jupiter::GFP FlyTrap line (CB05190).
Holland N., Holland D., Helentjaris T., Dhugga K.S., Xoconostle-Cazares B, & Delmer D.P. (2000). A Comparative Analysis of the Plant Cellulose Synthase (CesA) Gene Family. Plant Physiology, 123(4), 1313-1324.
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